Dataset Information

Role of LptD in Resistance to Glutaraldehyde and Pathogenicity in Riemerella anatipestifer.

ABSTRACT: Riemerella anatipestifer is a gram-negative bacterium that causes disease in ducks and other birds. Despite being an important pathogen in poultry, the pathogenesis and drug resistance mechanisms of this bacterium are poorly understood. An analysis of our unpublished RNA-Seq data showed that lptD, a gene encoding one of the lipopolysaccharide transport components, is transcribed at higher levels in strain CH-1 than in strain ATCC11845. In addition, strain CH-1 has been shown to display broader drug resistance than strain ATCC11845. Since LptD is involved in LPS biogenesis and drug resistance, we wondered if lptD is associated with increased R. anatipestifer resistance to glutaraldehyde, a disinfectant used in the production industry. In this study, the minimal inhibitory concentration (MIC) of glutaraldehyde for strain CH-1 was determined to be 0.125% (vol/vol), whereas an MIC of 0.05% (vol/vol) was observed for strain ATCC11845. Furthermore, the level of lptD transcription in strain CH-1 was consistently 2-fold higher than that observed in strain ATCC11845. Moreover, lptD transcription was upregulated in both strains at a subinhibitory concentration of glutaraldehyde. The role of lptD in R. anatipestifer was further assessed by constructing an ATCC11845 mutant strain with low lptD expression, R. anatipestifer ATCC11845 lptD -. The growth of R. anatipestifer ATCC11845 lptD - was severely impaired, and this strain was more susceptible than the wild-type strain to glutaraldehyde. Moreover, compared to the wild-type strain, R. anatipestifer ATCC11845 lptD - exhibited decreased biofilm formation and was more sensitive to duck serum. Finally, low lptD expression led to decreased colonization in ducklings. These results suggest that LptD is involved in R. anatipestifer glutaraldehyde resistance and pathogenicity.

SUBMITTER: Huang L

PROVIDER: S-EPMC6598057 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Role of LptD in Resistance to Glutaraldehyde and Pathogenicity in Riemerella anatipestifer.

Huang Li L Wang Mingshu M Mo Ting T Liu Mafeng M Biville Francis F Zhu Dekang D Jia Renyong R Chen Shun S Zhao Xinxin X Yang Qiao Q Wu Ying Y Zhang Shaqiu S Huang Juan J Tian Bin B Liu Yunya Y Zhang Ling L Yu Yanling Y Pan Leichang L Rehman Mujeeb Ur MU Chen Xiaoyue X Cheng Anchun A

Frontiers in microbiology 20190621

Riemerella anatipestifer is a gram-negative bacterium that causes disease in ducks and other birds. Despite being an important pathogen in poultry, the pathogenesis and drug resistance mechanisms of this bacterium are poorly understood. An analysis of our unpublished RNA-Seq data showed that lptD, a gene encoding one of the lipopolysaccharide transport components, is transcribed at higher levels in strain CH-1 than in strain ATCC11845. In addition, strain CH-1 has been shown to dis ...[more]

PMID: 31281307

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Project description:BackgroundRiemerella anatipestifer (R. anatipestifer) is one of the most important poultry pathogens worldwide, with associated infections causing significant economic losses. Rifampin Resistance is an important mechanism of drug resistance. However, there is no information about rpoB mutations conferring rifampin resistance and its fitness cost in Riemerella anatipestifer.ResultsComparative analysis of 18 R.anatipestifer rpoB sequences and the determination of rifampin minimum inhibitory concentrations showed that five point mutations, V382I, H491N, G502K, R494K and S539Y, were related to rifampin resistance. Five overexpression strains were constructed using site-directed mutagenesis to validate these sites. To investigate the origin and fitness costs of the rpoB mutations, 15 types of rpoB mutations were isolated from R. anatipestifer ATCC 11845 by using spontaneous mutation in which R494K was identical to the type of mutation detected in the isolates. The mutation frequency of the rpoB gene was calculated to be 10- 8. A total of 98.8% (247/250) of the obtained mutants were located in cluster I of the rifampin resistance-determining region of the rpoB gene. With the exception of D481Y, I537N and S539F, the rifampin minimum inhibitory concentrations of the remaining mutants were at least 64 μg/mL. The growth performance and competitive experiments of the mutant strains in vitro showed that H491D and 485::TAA exhibit growth delay and severely impaired fitness. Finally, the colonization abilities and sensitivities of the R494K and H491D mutants were investigated. The sensitivity of the two mutants to hydrogen peroxide (H2O2) and sodium nitroprusside (SNP) increased compared to the parental strain. The number of live colonies colonized by the two mutants in the duckling brain and trachea were lower than that of the parental strain within 24 h.ConclusionsMutations of rpoB gene in R. anatipestifer mediate rifampin resistance and result in fitness costs. And different single mutations confer different levels of fitness costs. Our study provides, to our knowledge, the first estimates of the fitness cost associated with the R. anatipestifer rifampin resistance in vitro and in vivo.

Dataset Information

Role of LptD in Resistance to Glutaraldehyde and Pathogenicity in Riemerella anatipestifer.

Publications

Role of LptD in Resistance to Glutaraldehyde and Pathogenicity in <i>Riemerella anatipestifer</i>.

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