Investigation of the replication capacity of lumpy skin disease virus in ticks

Ahmed #AbdelHaleem#ElWakil |Ausama #Abd El Raouf  #Yousif |Adel  #AbdelAzim#Fayed|Ibrahim  #Mohamed#elsabagh|Mahmoud  ##El Gamal|Omnia  #Hamdy#Refaei|

Abstract
Background: Lumpy skin disease (LSD) is a highly contagious trans- boundary disease of cattle with major economic losses. Lumpy skin disease virus (LSDV) belongs to the genus capripoxvirus of Poxviridae. Blood-feeding arthropods like mosquitoes, flies and, African tick species play a role in LSDV maintenance and transmission. Mechanical transmission of LSDV is well documented; however, evidence of transovarian and transstadial transmission is a tick warrants an investigation into the replication capacity of the virus in ticks.
Objective: Investigation of LSDV replication capacity in ticks using targeted detection of the viral DNA polymerase mRNA and metagnomics analysis using MinIon technology.
Methods: A total of 34 adult female ticks were collected from three LSD clinically disease cattle that were previously vaccinated using the Egyptian sheep pox (SPPV) vaccine, apparently healthy cattle in contact with the diseased cattle, and one healthy animal located in a previously infected zone. LSDV-specific PCR was used to detect LSDV in sampled ticks. PCR-positive tick samples were subject to a modified RT-PCR assay to detect the polyadenylated LSDV DNA polymerase mRNA.Metagenomic analysis using MinIon technology was used to generate LSDV sequences and to investigate relative abundance of LSDV sequences in the sampled tick population compared to other known organisms detected.
Results: Five ticks randomly selected form those collected were PCR positive for LSDV. A pool of 6 samples randomly selected from the collected ticks was also positive for LSDV using PCR. The modified RT PCR gave inconclusive results. Metagenomic analysis indicated the presence of a high genomic load of different known commensal and pathogenic microorganisms that have been reported to replicate in ticks (e.g. Staphylococci, streptococci and babesia spp.) In addition, the analysis revealed a LSDVsequence output that was nearly equal to sequence output of different bacteria known to replicate in ticks.
Conclusion: The modified RT-PCR assay used to detect the polyadenylated LSDV DNA polymerase mRNA in combination with data generated using metagenomic analysis provided additional support to the hypothesis that LSDV can replicate in ticks and, warrants further investigation into the replication capacity of LSDV in ticks.

Key words: LSDV; Metagenomic analysis; DNA polymerase; RT PCR, MinIon technology.

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