Abstract
Moraxella bovoculi has been isolated frequently from cattle with Infectious bovine keratoconjunctivitis (IBK). Two diverse genotypes of M. bovoculi, 1 and 2 were identified based on whole genome sequence analysis. It is essential to discriminate between the two genotypes to frame prevention and control measures. The whole genome of M. bovoculi TN7 was sequenced and compared to other M. bovoculi strains available in the NCBI database. M. bovoculi TN7 was found to be genotype 1, had an RTX toxin operon and pilA gene that are the known virulence factors in related Moraxella sp., but lacked antimicrobial resistance genes. M. bovoculi was found to have an open pangenome with 4051 (75.31%) accessory genes, and the addition of each new genome adds 18 genes to the pangenome. Comparison of pilin protein amino acid sequences revealed three new sequence types. Furthermore, the presence of linx, nagL, swrC and mdtA genes was found to be genotype 1 specific, whereas hyaD, garR, gbsA, yhdG, gabT, iclR, higB2, hmuU, hmuT and hemS were found only in genotype 2. Polymerase Chain Reaction (PCR) primers were designed and evaluated on strain TN7 plus seven additional strains accessible to us that had not been whole genome sequenced. This initial evaluation of the designed primers for the linX and hyaD genes produced the expected banding patterns on PCR gels for genotypes 1 and 2, respectively, among the 8 strains. The genotype-specific genes identified in this study can be used as markers for accurate diagnosis of genotype 1 isolates and this can aid in the development of autogenous or other molecular vaccines for treatment of infectious bovine keratoconjunctivitis (IBK) in resource-limited research settings.
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Data availability statement
Genome sequence of Moraxella bovoculi TN7 was submitted to NCBI. Corresponding accessions numbers with respect to BioProject, BioSample and SRA are as follows PRJNA792938, SAMN24477548 and SRR17373549.
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Acknowledgements
The authors are thankful to Tamil Nadu Veterinary and Animal Sciences University Chennai, India for providing necessary infrastructure to carry out the research work.
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KK: Conceptualized the work and carried out analysis, drafting the manuscript, SA: analysis of pangenome, MAC, PCR for genotyping and RS: overall monitoring of the work and fund acquisition for the work. All authors were involved in drafting and editing of the manuscript.
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Supplementary Fig. 1 Genome of Moraxella bovoculi TN7. The outer two circles demonstrate the coding sequence, tRNA, and rRNA of M. bovoculi TN7 annotated by Prokka pipeline. The third circle represents the GC skew curve (positive GC skew, green; negative GC skew, violet). The fourth circle shows the GC content (black) (JPG 4954 KB)
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Supplementary Fig. 2 Pangenome curve of M. bovoculi. A. Gene accumulation curves of the pangenome (blue) and core genome (green) of M. bovoculi. B. Number of new genes added with addition of each new M. bovoculi genome (JPG 2114 KB)
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Supplementary Fig. 3 Core genome phylogeny of M. bovoculi strains. Genotype 1 are marked in green color and genotype 2 in yellow color. Red color star marks on the phylogeny indicates 100% bootstrap value (JPG 3280 KB)
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Karthik, K., Anbazhagan, S., Chitra, M.A. et al. First report of the whole genome of Moraxella bovoculi genotype 1 from India and comparative genomics of Moraxella bovoculi to identify genotype-specific markers. Arch Microbiol 204, 663 (2022). https://doi.org/10.1007/s00203-022-03276-z
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DOI: https://doi.org/10.1007/s00203-022-03276-z