Molecular Characterization and Cytopathogenecity of Lumpy Skin Disease Virus in Egypt
Mohamed Mahmoud Mashaly;
Abstract
In recent LSD outbreaks during 2016-2019, there were many complaint from the cattle farms owners and breeders due to the presence of symptoms similar to those of LSD causing heavy losses so it necessitating highlighting, trying to characterize the cause of this complaint and the source of outbreaks that rapidly spread throughout the Egyptian governorates due to extreme importance to the owners of cattle farms and cattle industry for this purpose many points are investigated.
Lumpy skin disease outbreaks were observed during 2016 to 2019 in different governorates of Egypt affecting cattle and buffaloes throughout the year. The progressive shift of the outbreak in Egypt during 2018 causing severe losses in cattle population, the situation raising concerns that the disease continues to spread despite excessive vaccination campaigns.
During this study, many samples were collected from suspected animal from different governorates including (skin nodules and whole blood), an different assays were conducted on them to; confirm the presence of LSDV in suspected outbreak, isolation of the virus, molecular characterization and sequence analysis for the virus causing the recent outbreaks, in addition to study growth kinetic of Egyptian LSDV isolate in Vero and MDBK cell line.
In this study a total of 204 samples were examined by real-time PCR specific for detection of LSDV and 181 of the tested samples were positive for LSDV with different Ct values and total of 159 samples were positive with clear and obvious band and of predicted size of GPCR gene fragment 554bp
The detection of LSDV in three skin nodular samples of clinically suspected cattle by three blind passages on MDBK cell cultured. The isolated virus induced the characteristic CPE which appeared in the form of cell rounding, cell aggregation and cells coalesce together forming clusters and the CPE increased gradually till 70-90% of the sheet become completely detached.
Our trial for virus isolation of LSDV from skin biopsies (n=11) was successfully done on MDBK cell lines by three blind passages. Characteristic cytopathic effect (CPE) was observed in infected MDBK cells with suspected samples starting from the 5th day post inoculation in the first passage that become more prominent from the 3rd day in the second passage, The CPE was characterized by cell rounding with retraction of the cell membrane from surrounding cells and clustering to form rosette shape aggregations scattered all over the monolayer sheet starting from the 3rd day post inoculation which gradually increased leading to complete detachment of the cell monolayer at 7th day post inoculation.
Lumpy skin disease outbreaks were observed during 2016 to 2019 in different governorates of Egypt affecting cattle and buffaloes throughout the year. The progressive shift of the outbreak in Egypt during 2018 causing severe losses in cattle population, the situation raising concerns that the disease continues to spread despite excessive vaccination campaigns.
During this study, many samples were collected from suspected animal from different governorates including (skin nodules and whole blood), an different assays were conducted on them to; confirm the presence of LSDV in suspected outbreak, isolation of the virus, molecular characterization and sequence analysis for the virus causing the recent outbreaks, in addition to study growth kinetic of Egyptian LSDV isolate in Vero and MDBK cell line.
In this study a total of 204 samples were examined by real-time PCR specific for detection of LSDV and 181 of the tested samples were positive for LSDV with different Ct values and total of 159 samples were positive with clear and obvious band and of predicted size of GPCR gene fragment 554bp
The detection of LSDV in three skin nodular samples of clinically suspected cattle by three blind passages on MDBK cell cultured. The isolated virus induced the characteristic CPE which appeared in the form of cell rounding, cell aggregation and cells coalesce together forming clusters and the CPE increased gradually till 70-90% of the sheet become completely detached.
Our trial for virus isolation of LSDV from skin biopsies (n=11) was successfully done on MDBK cell lines by three blind passages. Characteristic cytopathic effect (CPE) was observed in infected MDBK cells with suspected samples starting from the 5th day post inoculation in the first passage that become more prominent from the 3rd day in the second passage, The CPE was characterized by cell rounding with retraction of the cell membrane from surrounding cells and clustering to form rosette shape aggregations scattered all over the monolayer sheet starting from the 3rd day post inoculation which gradually increased leading to complete detachment of the cell monolayer at 7th day post inoculation.
Other data
| Title | Molecular Characterization and Cytopathogenecity of Lumpy Skin Disease Virus in Egypt | Other Titles | التوصيف الجزيئى و التغيرات الخلوية المرضية لفيروس مرض الجلد العقدى فى مصر | Authors | Mohamed Mahmoud Mashaly | Issue Date | 2020 |
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