أحمد
أحمد

Ahmed

Assistant Professor of Bacteriology, Mycology and Immunology

Basic Informations

C.V


Curriculum Vitae

Ahmed Hussein Abed Moawad

(Ph.D." Microbiology", M.V.Sc." Microbiology", B. V. Sc.).

Bacteriology, Mycology and Immunology Departement, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef (62511), Egypt

Cell/ +2-0114-8824447; Work/ +2 082 2322066 / 117; FAX/+2 082 2327982.

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PERSONAL DETAILS:

Name

Ahmed Hussein Abed Moawad

Family name

Abed

Fore name

Ahmed

Date of birth

May 1, 1980

Place of birth

Beni-Suef, Egypt.

Nationality

Egyptian

Religion

Muslim

Marital status

Married, with two Children.

Correspondence address

Bacteriology, Mycology and Immunology Dept., Faculty of Veterinary Medicine, Beni-Seuf Univ, Beni-Suef, Egypt.

Tel. No.

Work:

FAX:

+201148824447

+2 082 2322066/ 117

+2 082 2327982

E-mail

 

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ACDADEMIC DETAILS - HIGHER EDUCATION

(Sep. 2003): Bachelor in Veterinary Medicine (B. V. Sc.)

Institution:           Faculty of Veterinary Medicine, Cairo University, Beni-Suef branch.

Consideration: The study extended from Sept.1998 to Sep.2003

General Grade: Very good with degree of honour   

Score:                 78.5%

(16 Oct. 2007): Master of Microbiology and Immunology, College of Veterinary Medicine, Beni-Suef University.

Institution:           Faculty of Veterinary Medicine, Beni-Suef University.

Thesis title: "A contribution towards the bacterial pathogens associated with respiratory problems in broiler chickens"

Supervised by: Prof. Fawzy R.El-Seedy and Prof. Ismail A. Radwan.

(27 June 2011): Ph.D. of Bacteriology, Mycology and Immunology

Institution:           Faculty of Veterinary Medicine, Beni-Suef University.

Thesis title "Recent and conventional methods for identification of Mycobacterium bovis in farm animals"

Supervised by: Prof. Fawzy R.El-Seedy, Prof. Ismail A. Radwan, Ass prof. Walid H. Hassan and Prof. Dr. Essam A. Nasr.

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Work experience – Position

(11 Jan.2004 – 11 Dec.2007):

Demonstrator (instructor) in Bacteriology, Mycology and Immunology department, Faculty of veterinary medicine, Beni-Suef University.

 (12 Dec.2007- 26 June 20011)

Assistant lecturer of Bacteriology, Mycology and Immunology, Faculty of veterinary medicine, Beni-Suef University.

 (July 2011 till now)

Lecturer of Bacteriology, Mycology and Immunology, Faculty of veterinary medicine, Beni-Suef University.

(Sept. 2011 till May 2012)

Lecturer of Microbiology in the Faculty of Nursing, Beni-Suef University.

(Sept. 2011 till Now)

Lecturer of Advanced Microbiology in the Faculty of Postgraduate Studies for Advanced Sciences, Beni-Suef University.

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Resposibilties:

Teaching:

  • Teaching Practical and Diagnostic Bacteriology, Mycology and Immunology to undergraduates.
  • Teaching Practical and Diagnostic Bacteriology, Mycology and Immunology to postgraduates.
  • Teaching Microbiology to 1st year students in the Faculty of Nursing, Beni-Suef University.
  • Teaching Advanced Microbiology to postgraduates in the Faculty of Postgraduate Studies for Advanced Sciences, Beni-Suef University.
  • Teaching and training summer courses of microbiology to undergraduate for 13 years.
  • Design & updating in collaboration with other teaching members in the department for the microbiology courses for under and postgraduates.
  • Supervision on 3 master theses which had been approved by the examining committee.
  • Supervision on 8 master theses and 3 doctoral theses.

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Research assistance:

  • Working in my master research project which is about" "A contribution towards the bacterial pathogens associated with respiratory problems in broiler chickens". Isolation of different bacterial isolates infecting broilers with emphasis mostly to E. coli and P. aeruginosa detecting their virulence factors and antibiogram.
  • Extraction and purification of Lipopolysaccharides (LPSs) from both organisms then analyzed and compared by SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE).
  • In the experimental infection to 21 day old chicken with E. coli and P. aeruginosa are compared with that of their LPSs which gave the same clinical signs and Postmortem lesions as well as mortalities. Specific hyperimmune sera against inactivated LPSs of E. coli and P. aeruginosa were prepared, detected and titrated serologically then, gave protection when inoculated with each specific organism.

 

  • Working in my Ph.D. research project which is about " Recent and conventional methods for identification of Mycobacterium bovis in farm animals". Screening of cattle and buffalo farms in different governorates for M. bovis by tuberculin test then, Postmortem examination and organ culture from positive reactors. Isolation and identification of mycobacterial isolates.
  • Field evaluation of serodiagnosis (ELISA) using unique and cocktail antigens specific for M. bovis compared to PCR with primers detecting specific insertion sequences for M. bovis to detect bovine tuberculosis in cattle and buffaloes in Egypt.

 

  • From the end of my undergraduate studies till now, I worked in the lab in a team-work with my professors and my colleagues in the college in many of their researches for example a research concerned with the role of house rat in the transmission of zoonotic bacteria with emphasis on Salmonella to human food.
  • Together with my professor, Dr. Ismail Radwan, I worked in his lab in a research concerned with the pathogenesis of C.perfringens and how to inhibit its toxin production by using B. subtilis culture as a probiotic. We worked both in lab and using a bird model.

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Technical and Field Skills:

  • Practical microbiology techniques such as bacterial isolation and purification. Morphological identification using different stains such as Gram's, Ziehl Neelsen, Loeffler's MB, Polychrome MB, Leishman's and Giemsa stains as well as special stains (capsular, flagellar and spore). Biochemical and serological tests for identification of bacteria.
  • Comprehensive knowledge of evidence-based microbiology practice in intended areas of specialty, including related equipment, techniques, practices and procedures

Molecular techniques

  • DNA and RNA manipulation.
  • PCR.
  • Gel electrophoresis.

Different serological techniques

-       ELISA (either using single or cocktail coating antigens).

-       AGPT

-       Slide and tube agglutination tests.

-       CFT

  • Laboratory diagnosis of some infectious diseases as Brucellosis in farm animals.
  • Field and laboratory supervision on poultry flocks in Beni-Suef and El-Fayoum Governorates.

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Computer skills:

  • Passing ICDL from ministry of high education (Windows, word, power point, excel, access, information technology and Internet).

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 Language skills:

  • Arabic: Fluent of both spoken and written "mother language"
  • English: Good command of English. TOEFL score (pBT 503)

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Conferences and Activities

  • Participated in the preparation of Beni-Suef Vet. Med. J. (the faculty journal) in 2006.
  • Participate in Quality Assurance and Accreditation Project (QAAP) in the faculty.

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Memberships and awards

  • Membership in Veterinary medicine Syndicate, Egypt (2003-present).
  • Membership in Beni-Suef School of Veterinary medicine Scientific Association, Beni-Suef, Egypt (2003).
  • Exceptional student award in the Education Day held at Beni-Suef University, Egypt, 2001

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Training courses

  • University teacher preparation course for 12 days at Beni-Suef University (28/11- 9/12/2010).
  • Human Development at Beni-Suef University (2008).
  • Active participation in the visit of Prof. Dr. Thomas Craig, professor at the University of Texas , USA, from 11-13/7/2016 Which included workshops on :
    • Scientific writing and publishing.
    • Parasites of Farm Animals.
    • Impact of parasitic diseases for the veterinarian.
    • Anthelmintic  resistance : what's old? what's now? what's up?
  • Program and course specification
  • Time arrangement and work pressures at Beni-Suef University (16-18/5/2006)
  • Enhancement of communication skills at Beni-Suef University (27-29/5/2006).
  • Enhancement of thinking skills at Beni-Suef University (24-26/6/2006).
  • Technology application on teaching at Beni-Suef University (3-6/8/2006).
  • Scientific research ethics at Beni-Suef University (12-14/4/2011).
  • Working Ethics of university professions at Beni-Suef University (26-28/4/2011).
  • Competitive Research Projects at Beni-Suef University (7-8/8/2016).
  • Teaching credit hours System at Beni-Suef University (14-15/8/2016).
  • International Publishing of Research at Beni-Suef University (17-18/8/2016).
  • Time and Meeting arrangement at Beni-Suef University (17-18/8/2016)
  • University Management at Beni-Suef University (21-22/8/2016)

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Editions

v  Participate, in collaboration with other teaching members in the department, in design and updating the Microbiology courses for under and postgraduates:

  1. Notes on Veterinary Microbiology.
  2. Notes on Bacteriology, Mycology and Immunology.
  3. An introduction to Practical Veterinary Microbiology.

v  Design the Microbiology  and Immunology course for 1st year student in the Faculty of Nursing, Beni-Suef University.

  1. "Notes on Medical Microbiology& Immunology"

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References

Prof. Dr. /Ismail Abd El- Hafeez Radwan Rahil

Professor and Head of Microbiology Department, Faculty of Veterinary Medicine Beni-Suef University, Egypt.

Phone: +2 082 2322066 / 117

FAX: +2 082 2327982

E-mail: Microbiologist111@yahoo.com

 

Prof. Dr. /F.R. El-Seedy.

Professor of Microbiology. Faculty of Veterinary Medicine, Beni-Suef University, Egypt.

Phone: +2 082 2322066 / 117

FAX: +2 082 2327982

E-mail: frseedy@bsu.edu.eg

 

Ass.Prof. / Walid Hamdi Hassan.

Assistant professor of microbiology, Faculty of Veterinary Medicine, Beni-Suef University, Egypt.

Phone: +2 082 2322066 / 117

FAX: +2 082 2327982

E-mail: dr_walidh@yahoo.com

 

Prof. Dr. Essam Amin Nasr

Professor and Head research of Bacterial Diagnostic Products Department (Tuberculosis),

Veterinary Serum and Vaccine Research Institute- Abbasia, Cairo

E-mail: essamnasr@yahoo.com

 

Ass.Prof./ Ahmed Sayed Abel-Moneim.

Professor of virology, Faculty of Veterinary Medicine, Beni-Suef University, Egypt.

Phone: +2 082 2322066

FAX: +2 082 2327982

E-mail: a_s_abdel_moneim @ yahoo.com

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LIST OF PUBLICATIONS

  1. El-Seedy, F. R.; Radwan, I. A.; Hassan, W. H.; Nasr, E. A.; Abed, A. H. and Moussa, I. M. I. (2013): The correlation between M. bovis isolation and ELISA using PPD-B and ESAT6- CFP10 mixture on the sera of tuberculin reactor cattle and buffaloes. J. Food, Agric. Environ., 11(1):489-494. (Impact factor: 0.3).
  2. El-Nesr, K. A.; Mahdy, E. A.; Hassan, W. H. and Abed, A. H. (2013): Bovine Neurobrucellosis: Pathological and Bacteriological studies. Beni-Suef Vet. Med. J., 22(1):58-63.
  3. Zeinhom, M. M. A.; Abed, A. H. and Hashem, K. S. (2013):  A contribution towards milk enzymes, somatic cell count and bacterial pathogens associated with subclinical mastitis cows milk. Assiut Vet. Med. J.; 59(138):38-48.
  4. Radwan, I. A.; Abed, A. H.; Abeer, M. R.; Ibrahim, M.A. and Abdallah, A. S. (2014): Effect of Thyme, Clove and Cinnamon Essential Oils on Candida albicans and Moulds Isolated from Different Sources. Am. J. Anim. Vet. Sci., 9(4): 303-314. DOI: 10.3844/ajavssp.2014.303-314. (Impact factor: 0.47).
  5. Amer, M. M.; Dahshan, A-H. M.; Abed, A. H. and Fahmy, S. S. (2016): Phenotypic and genotypic characterization of Clostridium perfringens associated with necrotic enteritis in broiler chickens. J. Vet. Med. Res., 23 (1):50-56. J. homepage: http://www.bsu.edu.eg/bsujournals/JVMR.aspx. Online ISSN: 2357-0520. Print ISSN: 2357-0512.
  6. El-Seedy, F. R.; Abed, A. H.; Yanni, H. A. and Abd El-Rahman, S. A. A. (2016): Prevalence of Salmonella and E. coli in neonatal diarrheic calves. Beni-Suef Univ. J. of Basic and Appl. Sci., 5:45-51. Available online at www.sciencedirect.com. J. homepage: www.elsevier.com/locate/bjbas.
  7. Osman, K. M.; Ali, M. N.; Radwan, I. A.; ElHofy, F.; Abed, A. H.; Orabi, A. and Fawzy, N. M. (2016): Dispersion of the vancomycin resistance genes vanA and vanC of Enterococcus isolated from Nile tilapia on retail sale: A public health hazard. J. Frontiers Microbiol., Volume (7) Article (1354). (Impact factor: 4.165).
  8. Radwan, I. A.; Abed, A. H., Abd Allah, M. M. and Abd El-Latif, M. A. A. (2016): Bacterial pathogens associated with cellulitis in chickens. J. Vet. Med. Res., (accepted for publication).
  9. Radwan, I. A.; Abed, A. H.; Abd Al-Wanis, S. A.; Abd El-Aziz, G. G. and El-Shemy. A. (2016): Antibacterial effect of cinnamon and oreganium oils on multidrug resistant Escherichia coli and Salmonellae isolated from broiler chickens. J. Egy. Vet. Med, Ass., 76(2):169-186.
  10. Radwan, I. A.; Abed, A. H. and Abd El-Aziz, M. M. (2016): Fungal pathogens associated with respiratory problems in broiler chickens. J. Vet. Med. Res., 23 (1):57-64. Journal homepage: http://www.bsu.edu.eg/bsujournals/JVMR.aspx. Online ISSN: 2357-0520. Print ISSN: 2357-0512.
Radwan, I. A.; Shehata, A. A. E.; Abed, A. H. and Hosni, A. R. (2016): Bacterial Species Associate Broiler Proventriculitis and their Antimicrobial Resistance. J. Vet. Med. Res., (accepted for publication).
C.V PDF

Master Title

A contribution towards the bacterial pathogens associated with respiratory problems in broiler chickens

Master Abstract

Bacteriological examination of a total of 300 chickens revealed that 235 cases were harbored bacterial strains affecting the respiratory tracts of chickens with an incidence of 78.33%. Out of 300 cases, 212 and 23 cases werepositive for single and mixed infection with an incidence of 70.67% and 7.67%, respectively. The bacterial species isolated from the examine cases (300) were predominantly, E. coli (125 isolates), P. aeruginosa (64 isolates), Kl. pneumoniae subsp. pneumoniae (29 isolates), P. mirabilis (16 isolates), S. aureus (14 isolates) and Streptococcus species (10 isolates). The most prevalent E. coli serogroups recovered from diseased chickens with respiratory manifestations were O78 and O1 with an incidence of 19%, 7.33%, respectively, followed by O2 (4%), O8 (3.33%), O25 (2.67%) and O119 (2%) while 10 isolates (3.33%) could not be serotyped by the available antisera. E. coli, P. aeruginosa, P. mirabilis and S. aureus were recovered mainly from air sac samples (27%, 10.67%, 3.67% and 2%, respectively). Meanwhile, Str. avium and Str. zooepidemicus were recovered mainly from lung samples (1.33% and 0.67%, respectively). Kl. pneumoniae subsp. pneumoniae were recovered mainly from both lung and tracheal samples (3.33% of each). The isolated E. coli serogroups were examined for the following virulence factors: Congo red binding assay, haemolytic activity, haemagglutination activities (haemagglutination and mannose resistance haemagglutination) and serum resistance tests (survival or growth in serum). The serogroups O78 and O1 were the most virulent isolates. Only serogroups O78 and O1 showed Congo red binding activity with an incidence of 68.7%. None of the isolates showed haemolytic activity while, all serogroups showed mannose resistance activity with chicken erythrocytes with an incidence of 100%. Also, all serogroups except serogroup O25 was able to survive and grow in serum with an incidence of 93.54% for each. Concerning with detection of the virulence of P. aeruginosa it was found that, 58 isolates (90.63%) were ß- haemolytic onto blood agar, while only 6 isolates (9.37%) were non haemolytic. All isolates produced exotoxins which have bactericidal effect on the other microorganisms. The in-vitro sensitivity of 20 isolates of E. coliO78, 15 isolates of E. coliO1and 25 isolates of P. aeruginosa against 13 chemotherapeutic agents revealed that, E. coli O78 isolates were highly sensitive to Gentamycin (90%), Florfenicol (85%) and LincoSpectin (75%). The same strains were highly resistant toAmoxycillin (75%), Erythromycin (70%) and Streptomycin (70%).E. coliO1 isolates were highly sensitive to Florfenicol, LincoSpectin and Gentamycin in an incidence of 86.67%, 86.67% and 80%, respectively. On the other hand, E. coliO1 strains were highly resistant to Amoxycillin (86.67%), followed by Streptomycin (66.67%) then Erythromycin and Kanamycin (60%) for each. P.aeruginosa strains were more sensitive to Enrofloxacin (40%), Pefloxacin (36%) and Ceftifour (36%).The results also showed that, P.aeruginosa strains were resistant to the most chemotherapeutic agents especially Amoxycillin (100%), Erythromycin (80%) and Kanamycin (76%). Lipopolysaccharides (LPSs) were extracted and purified from the cell wall of E. coliO78 and P.aeruginosa then analyzed by Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS- PAGE) and stained with silver stain. The results revealed that, presence of similarity between LPS of E. coliO78 and that of P.aeruginosa. Hyperimmune sera were prepared in chickens against inactivated LPSs of both E. coliO78 and P.aeruginosa. The obtained hyperimmune sera were examined for detection and titration of specific antibodies against specific LPS using AGPT and ELISA and against the specific microorganisms using slide agglutination test. Results of AGPT revealed that, each hyperimmune serum gave positive results with the both LPSs of E. coliO78 and P.aeruginosa. Results of slide agglutination tests for chicken hyperimmune sera against the whole cells of E. coli and P. aeruginosa showed that, each hyperimmune serum gave positive agglutination with its specific microorganism. Results of ELISA showed that, the antibody titer of E. coli LPS was 800 and the antibody titer of P.aeruginosaLPS was 400. The experimental infections of isolated E. coliO 78 and P. aeruginosa and their LPSs for 21 days old chicks either through air sacs or intravenously were carried out and compared with each other and the results showed similarity between each microorganism and its specific LPS in clinical signs, post mortem pictures an mortalities. Each microorganism was inoculated with the specific hyperimmune serum against its LPS. It was found that each hyperimmune serum gave protection against its specific microorganism when inoculated in chickens.

Master Abstract PDF

PHD Title

Recent and conventional methods for identification of Mycobacterium bovis in farm animals

PHD Abstract

Bovine tuberculosis is a chronic bacterial disease caused by M. bovis. It can infect many species of animals; cattle and buffaloes most commonly and are considered the main¬tenance hosts for the bacteria. Bovine tuberculosis can spread to humans. It is still common in developing countries, a source of economic loss, and a serious health threat to humans. The result recorded in this study illustrated the prevalence of tuberculin reactors in dairy cattle and buffaloes in different governorates. From a total of 3600 tuberculin tested cross-bred dairy cattle including 32 herds as follows: (El-Fayoum 450 animals, Alexandria road El-Sahrawy 1850 animals, Gharbia 550 animals and Beheira 750 animals), 72 were found to be reactors with a prevalence rate of 2%. The highest herd prevalence was present in Gharbia 60% and the lowest was present El-Sahrawy, 33.3%, while it was 40% in El-Fayoum and 57.1% in Beheira. The highest result for tuberculin test was 2.7% in Gharbia and the lowest result was 1.6% in El-Sahrawy. In Beheira, it was 2.3%, and El-Fayoum, it was 2.2%. Also, from a total of 2550 tuberculin tested buffaloes as follow (Beheira 500 animals, Gharbia 600 animals and Alexandria road El-Sahrawy 1450 animals), 26 were found to be reactors with a prevalence rate of 1%. The highest result for tuberculin test was 1.2% in Gharbia and the lowest result was 0.8% in Beheira, while in Cairo-Alexandria El-Sahrawy road, it was 1%. The relationship between the reactivity of tested cattle and buffaloes to tuberculin test and postmortem findings was investigated in this study. In cattle, it revealed that the overall percent of PM findings with VL was 49 out of 72 (68.1%), and the percent of tuberculin reactors with NVL was 23 out of 72(31.9%). While, in buffaloes, VL was 17 out of 26 (65.4%), and the NVL was 9 out of 23 (34.6%). Regarding the relationship between tuberculin reactors (cattle and buffaloes) and site of lesions; the 49 slaughtered tuberculin reactor cattle with VL were distributed in head 8 (11.1%), pulmonary 25 (34.7%), digestive 7 (9.7%), mixed 4 (5.6 %) and generalized 5 (6.9%). While in buffaloes, the 17 VL were distributed in pulmonary 3 (11.5%), digestive 7 (26.9%), mixed 6 (23.1 %) and generalized 1 (3.8%). Concerning the correlation between PM finding in different age of tuberculin reactor cattle; the number of tuberculin positive reactors cattle were 12 (1.2%) from 975 animals at age from 1-3 years; 5 (41.7%) showed VL and 7 (58.3%) showed NVL. At age from 3-5 years, the number of tuberculin positive reactor cattle were 39 (2.1%) from 1900 animals; 32 (82.1%) showed VL and 7 (17.9%) showed NVL while at age over 5 years, the number of tuberculin positive reactors were 21 (2.9%) from 725; 12 (57.1%) showed VL and 9 (42.9%) showed NVL. The total isolation rates of mycobacteria from carcasses of reactor cattle and buffaloes with and without lesions were summed up in this study. In cattle, from a total of 72 carcasses, 44 were positive cultures with an isolation rate of 61.1%, all isolates were identified as M. bovis; of them, 40 (81.6%) were from VL and 4 (17.4%) were from NVL. While in buffaloes, from a total of 26 carcasses, 15 were positive cultures with an isolation rate of 57.7%; 10 isolates (38.5%) were M .bovis, all were from the 17 VL (58.8%), and 5 isolates (19.2%) were MOTT, 3 (17.6%) were from the 17 VL and 2 (22.2%) were from the 9 NVL. Concerning the microscopical examination by ZN staining of the 72 specimens from the slaughtered reactor cattle showed the AFB in only 26 (36.1%) samples. While in buffaloes, the 26 specimens showed the AFB in 7 only (26.9%). On the other hand, the PM lesions (VL and NVL) of the 72 the slaughtered tuberculin reactor cattle were sorted into 4 degree of pathological lesions. Degree 0 represented all the 23 NVL (31.9%). On the other hand, the 49 VL were distributed as 3 Degree 1 (4.2%) and 23 (31.9%) for each of Degree 2 and Degree 3 which were the most predominant. Concerning the correlation between M. bovis isolation and degree of pathological lesions, out of the 72 slaughtered reactor cattle, a total of 44 M. bovis isolates were recoverd with a rate of isolation of 61.1%. Of them, 4 isolates (5.6%) were from Degree 0, 19 isolates (26.4%) were from Degree 2 and 21 isolates (29.2%) from Degree 3. No isolates were recovered from Degree 1. Regarding the comparison between results of tuberculin test and ELISA technique using PPD-B and ESAT6-CFP10 Mixture in cattle; out of 72 tuberculin reactors 49 (68.1%) serum samples were positive for ELISA using PPD-B and 45 (62.5%) serum samples were positive for ELISA using antigen mixture as coating antigens. And out of 49 tuberculin positive reactors with VL, 44 (89.8%) were positive for ELISA using PPD-B and 41(83.7%) by using antigen mixture and out of 23 tuberculin reactors with NVL, 5 (21.7%) were positive for ELISA using PPD-B, 4 (17.4%) were positive for ELISA using antigen mixture. Moreover, regarding the correlation between M. bovis isolation and ELISA using PPD-B and antigen mixture on the sera of tuberculin reactors cattle; out of 8 tuberculin reactor cattle with head lesions, 5 (62.5%) gave M. bovisisolates while 7 (87.5%) and 5 (62.5%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Out of 22 reactors with pulmonary lesions, 22 (88%) gave M. bovisisolates while 22 (88%) and 23 (92%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Out of 7 reactors with digestive lesions, 5 (71.4%) yielded M. bovisisolates, while 6 (85.7%) and 4 (57.1%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Out of 4 reactors with mixed TB lesions, 3 (75%) gave M. bovisisolates while all of them (100%) were ELSIA positive by using both PPD-B and antigen mixture. Out of 5 reactors with generalized TB lesions, all of them (100%) yielded M. bovisisolates and were ELSIA positive by using both PPD-B and antigen mixture. Out of 23 reactors with NVL, 4 (17.4%) gave M. bovisisolates while 5 (21.7%) and 4 (17.4%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Regarding the comparison between results of tuberculin test and ELISA technique using PPD-B and antigen mixture in buffaloes; out of 26 tuberculin reactor buffaloes, 16 (61.5%) serum samples were positive for ELISA using PPD-B, while 14 (53.8%) serum samples were positive for ELISA using antigen mixture. From other side, out of 17 tuberculin positive reactors with VL, 13 (76.5%) were positive for ELISA using PPD-B and 12 (70.6%) by using antigen mixture. On the other hand, out of 9 tuberculin reactors with NVL, 3 (33.3%) were positive for ELISA using PPD-B, 2 (22.2%) were positive for ELISA using antigen mixture. Moreover, regarding the correlation between M. bovisisolation and ELISA using PPD-B and antigen mixture on the sera of tuberculin reactors cattle; out of 3 tuberculin reactors buffaloes with pulmonary lesions, 2 (66.7%) gave M. bovisisolates and also were ELSIA positive by using both PPD-B and antigen mixture. Out of 7 reactors with digestive lesions, 4 (57.1%) gave M. bovisisolates while 6 (85.7%) and 5 (71.4%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Out of 6 reactors with mixed TB lesions, 3 (50%) gave M. bovisisolates while 4 of them (66.7%) were ELSIA positive by using both PPD-B and antigen mixture. The one reactor with generalized TB lesions gave one M. bovisisolate and was ELSIA positive by using both PPD-B and antigen mixture (100%). Out of 9 reactors with NVL, no M. bovisisolates were recovered (0%) while 3 (33.3%) and 2 (22.2%) were ELSIA positive by using PPD-B and antigen mixture, respectively. Regarding the sensitivity and specificity of ELISA using PPD-B in cattle evaluated in this study revealed 89.8% and 78.3%, respectively. On the other hand, the sensitivity and specificity of ELISA using antigen mixture revealed 83.7% and 82.6%, respectively. Concerning the buffaloes, the sensitivity and specificity of ELISA using PPD-B revealed 76.5% and 66.7%, respectively. On the other hand, the sensitivity and specificity of ELISA using antigen mixture revealed 70.6% and 77.8%, respectively. PCR test was applied on randomly selected 16 mycobacterial isolates, which are physically and biochemically identified as 14 M. bovis and 2 MOTT. Of the 14 M. bovis isolates, 10 were from cattle distributed as one head, 3 pulmonary, 2 digestive, one mixed, 2 generalized forms and one NVL) and 4 from buffaloes (one from each of pulmonary, digestive, mixed and generalized forms). On the other hand, the 2 MOTT were from buffaloes (one from each of digestive and NVL). The results revealed that all the 14 isolates confirmed physically and biochemically as being M. bovis gave positive results with PCR test using Oligonucleotide primer that amplifies a 350bp fragment in RD7 region of M. bovis (100%) and the 2 MOTT isolates gave negative results with the PCR test by using the same primer providing confirmation that it did not belong to the M. bovis as they lake the a target for primers. Recommendations ? It is highly recommended that the application of "test and slaughter" policy is an essential mean to prevent spreading of this serious disease among cattle and buffalo herds in Egypt. ? Testing of new introduced animals at purchase to identify positive animals and avoiding purchasing old animals from markets might help decreasing within- herd prevalence. ? Good hygiene, housing and management reduce the prevalence of bovine tuberculosis. ? ELISA utilizingantigen mixture could detect early infection and also the advanced stages of infection as it counter the variability observed in antigen recognition throughout the infection process and it can be used complementary to the skin test todetermine the status of disease and reduce the frequency ofmisdiagnosis. ? The use of ELISA is suggested for situations where the investigation of the whole herd is more important than the individual testing of each animal. In addition, the ELISA can also be helpful when a collective diagnosis is desired to elucidate clinical suspicions of disease, or in the first steps of a control program, for identification of foci. ? The PCR technique can be used as a rapid confirmatory test. It is much faster than culture, reducing the time for diagnosis to 2 days and providing the ability to detect the presence of M. bovis DNA. Although direct PCR can produce a rapid result, it is recommended that culture be used in parallel to confirm the existence of a viable M. bovis in the positive reactor animals.

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