1- Department of internal medicine and infectious diseases, fac. of vet. Med. Menofia university- Sadat branch, Egypt

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Sero-diagnosis of bovine tuberculosis by ELISA using bovine PPD and ST.CF. ElSify A1.,Nayel M1., Hazem S2., Tarabess R3., Akram S1., Allaam M1., Hassan

H1., and El Garhy M2.

mohamed.aboelezz@vet.menofia.edu.eg

2- Animal Reproduction Research Institute, Agriculture Research Centre. AlHaram- Egypt

3- Department of bacteriology, mycology and immunology, fac. of vet. Med. Menofia university- Sadat branch, Egypt

Abstract.

Bovine tuberculosis represents one of the very important infectious diseases in Egypt and the world. It has zoonotic importance and causes severe economic losses. Accurate and rapid diagnosis considered as the milestone for control of the disease. In this study ELISA technique was used for confirmation of positive reactors cows that tested with single intradermal tuberculin test, to detect false positive reactors. Bovine PPD and ST.CF antigens have been used as two different coating antigens for ELISA technique. 3747 cattle from dairy farms in five different governorates were subjected to the single intradermal cervical tuberculin test whereas 78 (2.24%) proved positive reactors to tuberculin. These positive reactors tested with ELISA. 64 (82.05%) animals were positive by

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ELISA coated with ST-CF, while by using bovine PPD as coating antigen 58 (74.35%) animals were positive. The previous results indicated that ELISA test showed higher sensitivity and specificity using ST-CF as coating antigen than in case of bovine PPD coating antigen.

Key words:

Bovine tuberculosis, Mycobacterium bovis, Diagnosis, ELISA, PPD and ST.CF.

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Molecular typing of Clostridium perfringens isolates from soil, healthy, and diseased sheep in Egypt by multiplex PCR.

Nayel M1., ElSify A1 ., Akram S 1., Allaam M1., Eman Abdeen2 and Hassan H1.

1- Department of internal medicine and infectious diseases, fac. of vet. Med. Menofia university- Sadat branch, Egypt

2- Department of bacteriology, mycology and immunology, fac. of vet. Med. Menofia university- Sadat branch, Egypt

Abstract.

In this study we use the multiplex PCR for typing of isolates of Clostridial perfringens from soil, clinically healthy, diseased sheep. Clostridium perfringens was isolated from 41 of 100 soil samples, from 12 of 100 clinically healthy sheep and from 118 of 200 sheep with enterotoxaemia signs. Genotyping of 41 isolates from soil indicated that 29 (70.73%) were type A, 3 (7.31%) were type B and 9 (21.95%) were type D. Of 12 isolates from clinically healthy sheep indicated that 6 (50%) were type A and 6 (50%) were type D. Of 118 isolates from diseased sheep indicated that 42 (35.59%) were type A, 22 (18.64%) were type B and 54 (45.76%) were type D. This indicated that Clostridium perfringens type A, B and D are the main types causing diseases in sheep in Egypt and Clostridium perfringens type A must be included in any vaccine programme to ensure optimum protection.

Key words:Sheep clostridial enterotoxaemia, multiplex PCR and Clostridium

perfringens.

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The use of different diagnostic tools for Babesia and Theileria

parasites in cattle in Menofia, Egypt

Mohamed Nayel1, Khaled Mohamed El-Dakhly2, Mahmoud Aboulaila3, Ahmed Elsify1, Hany Hassan 1, Elsayed Ibrahim1, Akram Salama1, Tokuma Yanai4.

1-Department of internal medicine and infectious diseases, fac. of vet. Med. Menofia university- Sadat branch, Egypt

3- Department of Parasitology, fac. of vet. Med. Menofia university- Sadat branch, Egypt

Abstract:

Bovine piroplasmosis is caused by tick-borne hemoprotozoans of the genera Babesia and Theileria and is the most prevalent in tropical and subtropical countries, causing a major economic impact worldwide. In the current study, a total of 405 cattle of different ages, sexes, and breeds were randomly sampled for surveying and diagnosis of babesiosis and theileriosis using three methods: direct microscopy (blood smears), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Giemsa stained blood smears revealed that, out of 405 examined cattle, 33 (8.15 %) were infected with Babesia sp. and 65 (16.05 %) with Theileria sp. (total number of infected cattle was 98). Mixed infection was seen in 11 (2.72 %) animals. Moreover, application of the three diagnostic assays on 158 randomly sampled cattle indicated that 17 (10.76 %) and 33

(20.89 %) were positive for Babesia and Theileria spp. By the direct smear technique, 25 (15.82 %) and 33 (20.89 %) by IFAT (fluorescence was greenish yellow for Babesia and yellowish for Theileria), and 20 (12.66 %) and 38 (24.05 %) by PCR. Using primers specific for Babesia and Theileria spp., we found that diagnostic bands appeared at ~350 and ~370 bp, respectively indicating the presence of these piroplasms. Statistically, there was a non-significant difference of the positivity in response to the three techniques; thus, any of these methods can be described as useful for diagnosing blood parasites in both domesticated animals and birds. On the basis of

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the obtained results, it could be concluded that direct microscopy can be used in acute infections, whereas IFAT and PCR are useful in chronicity.

Key words: Blood parasites, Cattle, Infectous Published in: 2012

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Application of molecular techniques on equine herpes virus 1 infection

Nayel Mohamed1 , Hassan Hany1 and Hideto Fukushi2

1Department of internal medicine and infectious diseases, fac. of vet. Med. Menofia university- Sadat branch, Egypt

Abstract:

Based on Equine herpesvirus 1 (EHV-1) bacterial artificial chromosome clone of a neuropathogenic strain Ab4p, ORF76 encoding EUS9 was replaced with a selection cassette, rpsL-neo gene, to produce an ORF76 deletion mutant, Ab4p_∆ORF76. Transfection of RK-13 cells with Ab4p_∆ORF76 genome DNA produced infectious virus, indicating ORF76 is a nonessential gene. Deletion of ORF76 had no effect on the transcript expression of neighboring genes, ORF75 and ORF67. In a CBA/N1 mouse infection model following intranasal inoculation, the ORF76 revertant mutant of EHV-1 displayed a wild-type level of neurovirulence and neural spread in the olfactory pathway, but Ab4p_∆ORF76 was avirulent and failed to invade the CNS. In the olfactory mucosa containing the olfactory receptor neurons, Ab4p_∆ORF76 replicated with efficiency similar to that of the ORF76 revertant mutant of EHV-1. However, Ab4p_∆ORF76 was not transported to the olfactory bulbs. The growth of Ab4p_∆ORF76 in cultivated neural cells was one order of magnitude lower than that of parental and revertant viruses. These results indicated that the ORF76 (EUS9) is essential for the anterograde spread of the virus from the olfactory mucosa to the olfactory bulb in the mouse.

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Published In: 2010.

References

Allen, G.P. (2000) Equine rhinopneumonitis. In: OIE Manual of Standards for Diagnostic Tests

and Vaccines, 4th edn. Eds: M. Truszeynski, J.E. Pearson, S. Edwards and B. Schmitt. OIE Press, Paris, pp. 565-575.

Allen, G. P. (2002) Epidemic disease caused by Equine herpesvirus-1: recommendations for

prevention and control. EQUINE VETERINARY EDUCATION / AE / JUNE 2002, pp. 177-184.

Allen, G. P. (2006) New insights into equine herpesvirus-1 (EHV-1) neurologic disease. Equine

Disease Quarterly, 15, pp. 2-3.

Allen, G.P., Bolin, D.C., Bryant, U., Carter, C.N., Giles, R.C., Harrison, L.R., Hong, C.B., Jackson, C.B., Poonacha, K., Wharton, R. and Williams, N.M. (2008) Prevalence of latent,

neuropathogenic equine herpesvirus-1 in the Thoroughbred broodmare population of central Kentucky. Equine Vet. J. 40, pp. 105–110.

Allen, G.P. and Breathnach, C.C. (2006) Quantification by real-time PCR of the magnitude

and duration of leucocyte-associated viraemia in horses infected with neuropathogenic vs. non-neuropathogenic strains of EHV-1. Equine Veterinary Journal, 38, pp. 252-257.

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Serosurvey of Bovine Ephemeral Fever in Menoufia

Governorate

Mohamed Nayel1, Hany Hassan1 and Ahmed Zaghawa1

1Department of internal medicine and infectious diseases, fac. of vet. Med. Menofia university- Sadat branch, Egypt

In the present study, a total of 3700 Ferisian cattle and 120 local breed cattle of different ages and sex from Menoufia governorate were investigated clinically and serologically using SNT, NPLA and CBIA for BEF virus infection. A total of 1446 (39.08%) Ferisian cattle and 34 (28.33%) local breed cattle showed the clinical signs with case fatality 6.92% and 0% for Ferisian and local breed respectively. Animals of two- < three years showed the highest

prevalence of the disease in both Ferisian (39.90%) and local breed (37.87%). Isolation of BEF virus from leucocytic fractions of feverish cattle by blind intracerebral inoculation in baby mice was confirmed by positive Immunoperoxidase on fixed preparations of infected mice brain. Histopathological examination of infected mice brain showed the pathological changes of the brain tissue which explained the nervous manifestations observed on inoculated baby mice. BEF virus was also isolated on VERO cells which were inoculated by reconstituted suspension of infected mice brain. The isolation on tissue culture was confirmed by observation of specific cytopathic effect and by PLA performed on infected tissue culture and finally electron microscopy which showed the virus particles in the cytoplasm of infected VERO cells.

Preparation of hyper immune serum against isolated BEF virus resulted in good antisera of high titer.

Keywords: Bovine Ephemeral Fever, Serosurvey, Isolation

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Abu EI-Zein, E.M.; Gameel, A.A.; AI-Afaleq, A.L; A1-Gundi, O.A.I. and Bukhari, A. (1997): Bovine ephemeral fever in Saudi Arabia. Vet. Rec. 140, (24): 630-631

Abu EI-Zein, E.M.; Gameel, A.A.; AI-Afaleq, A.L; A1-Gundi, O.; A1-Bashier, A.M.; Zeedan, A.; A1-Mageed, H.A. and Abu Khadra, H. (1999): Observations on the recent

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Al-Gaabary, M.H.; Osman,S.A.; Foad,F.(2005): Comparative clinical and epidemiological

studies on bovine ephemeral fever in Sakha farm, Kafer El-Sheikh Egypt. Assuit vet. Med. J. 51 (104): 76-85