Isolation and Identification of Foot and Mouth Disease Sat-2 Virus Isolate During Egyptian Outbreak 2012

During April and May 2012, six additional outbreaks of FMD type SAT 2 were reported in Egyptian governorates: EL- Gharbiya (Kafr Qeretna, Al – Mahala Alkobra, Ebshaway Almalak, Qutoor) Governorate, extending to local cattle, buffaloes and dairy farms in Kafr El-Shaykh (Alsalmya, Fouah), AL - Minya (Village 3, Samalout, Mansafees, Abo Querkas), Dumyat (Ezbet 20, Kafr saad), ALexandria (Iraq Village, Al Aamerya), EL minufya (Monshaat Gerges, Ashmoun, Shatanoof, Ezbet Algalayla, Berket Alsabaa, Mahalet Sabak),Luxor (Al Gorna) and Al- Sharqiya (Al -Telleen, Menya Al kamh) Governorates. Suspected FMD virus samples (22 Tongue epithelium (TE), 12 vesicular fluid samples (VF)), in addition to, 10 cell culture grown virus of tongue epithelium origin, 6 cell culture grown virus of vesicular fluid origin, 3 positive control (O), 3 positive control (A) and 3 positive control (SAT2) were isolated on primary bovine kidney cells and Baby Hamster kidney cells and inoculated in baby mice. The assays used in this study involved CFT for typing of virus samples; 3ABC-FMD for differentiation between vaccinated and infected animals in any revealed protective immune response in the examined sera from Egypt 2012 outbreak and Indirect ELISA typing kits for typing of FMDV. PCR as an advanced confirmatory technique was done before submitting FMDV suspected clinical samples for Reference Laboratory of Foot and Mouth Disease in Pirbright, United Kingdom. Samples were isolated on primary bovine kidney and Baby Hamster kidney cell cultures, where all samples gave cytopathic effect. Also, all virus samples inoculated in suckling baby mice showed pathognomonic paralysis effect of the virus in hind limbs of mice, whereas the three negative controls failed to give any changes in cell culture or any paralysis in the hind limbs of mice. Serum samples collected from suspected infected cattle were examined using FMD-3ABC. All positive samples to 3ABC antibodies indicated that the animals were infected by FMDV. FMD antibody detection revealed infection of 62% of sera collected from Al- Gharbiyam, 100% from Kafr El-Shaykh, 61% from Dumyat, 37.5% from Alexandria, 45.5% from Al-Monufya, 72.7% from Luxor and 66.6% from Al-Sharqiya. The obtained results of 3ABC antibodies were subsequently confirmed by Indirect ELISA Kit, which revealed that the causative agent of the outbreak was FMDV SAT2. The prefinal confirmatory technique was polymerase chain reaction (PCR), which was done to detect clinically suspicious samples and differentiate FMDV. For official conformation, the isolated viruses had been sent to the Reference Laboratory for Foot and Mouth Disease in Pirbright, United Kingdom. Isolation of the causative agent is the first step for diagnosis of the disease. The first steps done on the samples collected (tongue epithelium and vesicular fluid) from the field during the last outbreak of FMD in Egypt at 2012 was isolation on primary bovine kidney, Baby Hamster kidney cell culture and Swiss baby mice. Secondary, CFT was used to detect and identify the serotype. Also, Indirect ELISA kits were used for virus typing. PCR was used as advanced technique for confirmation of the causative agent using target primers. Finally, the new outbreak of FMD in Egypt 2012 was due to FMD serotype SAT2.