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.
摘要:
经常从患有传染性牛角膜结膜炎(IBK)的牛中分离出博食莫拉氏菌。两种不同基因型的博沃武利分枝杆菌,基于全基因组序列分析鉴定1和2。区分两种基因型以制定预防和控制措施至关重要。对博沃武利分枝杆菌TN7的整个基因组进行了测序,并与NCBI数据库中可用的其他博沃武利分枝杆菌菌株进行了比较。bovoculiTN7被发现是基因型1,具有RTX毒素操纵子和pilA基因,这是相关莫拉氏菌中已知的毒力因子。,但缺乏抗菌素抗性基因。发现博沃武利M.有一个开放的pangenome,有4051个(75.31%)附属基因,每个新基因组的添加为pangenome添加了18个基因。菌毛蛋白氨基酸序列的比较揭示了三种新的序列类型。此外,Linx的存在,nagL,swrC和mdtA基因被发现是基因型1特异性的,而hyaD,garr,GBSA,yhdG,gabT,iclR,higB2,hmuU,hmuT和hemS仅在基因型2中发现。设计聚合酶链反应(PCR)引物,并在菌株TN7加上我们尚未全基因组测序的其他7个菌株上进行评估。对linX和hyaD基因的设计引物的初步评估分别在基因型1和2的PCR凝胶上产生了预期的条带模式,在8个菌株中。本研究中鉴定的基因型特异性基因可用作准确诊断基因型1分离株的标记,这可有助于在资源有限的研究环境中开发用于治疗感染性牛角膜结膜炎(IBK)的自体或其他分子疫苗。