Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken

Barbosa, Fernanda de Oliveira; Neto, Oliveiro Caetano de Freitas; Batista, Diego Felipe Alves; Almeida, A. Bugalho de; Rubio, Marcela da Silva; Alves, Lucas Bocchini Rodrigues; Vasconcelos, Rosemeri de Oliveira; Barrow, Paul; Berchieri, Ângelo

doi:10.1016/j.bjm.2017.01.012

Cited by 32 publications

(23 citation statements)

References 18 publications

(29 reference statements)

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“…Property of motility that helps, to either invade or evade the infection site is a useful property during the colonization and pathogenicity [28]. The observed swarming motility was different among the isolates.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Salmonella Dublin isolated from bovine and human hosts

et al. 2019

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Background Salmonella enterica subsp. enterica serovar Dublin (S. Dublin), a cattle adapted serovar causes enteritis, and systemic disease in bovines. The invasive index of this serovar far exceeds that of the other serovars and human infections often present as fatal or highly resistant infections. In this, observational study, phenotypic properties of human and bovine-derived isolates of S. Dublin along with antibiogram of common antimicrobials were evaluated. The multiplex PCR confirmed isolates were genotyped using 7-gene legacy MLST. MIC assay was done by broth microdilution method. Previously published protocols were used to assess the motility, biofilm formation and morphotype. Vi antigen was agglutinated using commercial antiserum. Caenorhabditis elegans infection model was used to evaluate the virulence potiential. Phenotyping experiments were done in duplicates while virulence assay was done in triplicates. Whole-genome sequencing was used to predict the genes responsible for acquired resistance and a genotype-phenotype comparison was made. Results We evaluated 96 bovine and 10 human isolates in this study. All the isolates belonged to ST10 in eBG53 and were negative for Vi-antigen. The swarming motility, biofilm formation and morphotype were variable in the isolates of both groups. Resistance to sulfamethoxazole, ampicillin, chloramphenicol, tetracycline was > 90% in animal isolates whereas resistance to sulfamethoxazole was > 70% in human isolates. MDR was also higher in animal isolates. Human isolates were significantly (P < 0.0001) more virulent than animal isolates on C. elegans infection model. The genomic comparison based on the core SNPs showed a high degree of homogeneity between the isolates. The carriage of IncA/C2 plasmid was seen as a typical feature of isolates from the bovine hosts. Conclusion Human isolates showed more diversity in the phenotypic assays. Animal isolates showed a higher degree of antimicrobial resistance with greater MDR but human isolates formed more biofilm and had greater swarming motility as well as increased virulence to the nematode C. elegans. The carriage of IncA/C2 plasmid could contribute to the distinguishing feature of the bovine isolates. The tandem use of genotypic-phenotypic assays improves the understanding of diversity and differential behaviour of the same serovar from unrelated host sources.

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Section: Discussionmentioning

confidence: 99%

Characterization of Salmonella Dublin isolated from bovine and human hosts

et al. 2019

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“…The major function of flagellum is to enable organism possess swimming and swarming motility. Furthermore, flagellum has been demonstrated to act as a virulence factor such as adhesion, invasion, biofilm formation of the pathogen (Parker and Guard-Petter 2001; Duan et al 2013; Chaban et al 2015; Barbosa et al 2017). In addition, the flagellin (filament subunit protein) was considered as a potential target candidate for Salmonella detection, since the protein is abundantly expressed, located on the surface of bacteria, and has good antigenicity (Rumbo et al 2006; Mizel and Bates 2010; Hiriart et al 2013; Hajam et al 2017).…”

Section: Introductionmentioning

confidence: 99%

A promising detection candidate for flagellated Salmonella spp.

et al. 2019

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Salmonella is a common and important pathogen for both human and animals. All Salmonella except Salmonella pullorum and Salmonella gallinarum have flagellum. Flagellin (FliC) is the main subunit protein forming the bacterial filament, which is present in large amounts on the surface of all flagellated Salmonella . After bioinformatics analysis, the most highly conserved region (locates position from 1 to 102 amino acid residue of FliC, we named it as FliC′) was selected, and corresponding recombinant FliC′ (rFliC′) protein was tailored as an immunogen to generate monoclonal antibodies (MAbs) against Salmonella flagellin. BALB/c mice were immunized with the purified recombinant protein rFliC′, which were prepared by prokaryotic expression system pET22b (+) expressing FliC′. After fusion of spleen cells from the immunized mice and SP2/0 cells, three hybridoma cells (1D6, 2G6 and 3E2) producing MAbs against targeted flagellate Salmonella FliC′ were generated and screened. The ability of MAb 3E2 to recognize and bind to Salmonella flagella was demonstrated by immunogold electron microscopy (IEM) method. Western blot (WB) analysis demonstrated that MAb 3E2 could specifically recognize flagellated Salmonella strains. Moreover, MAb 3E2 has a direct agglutination activity against Salmonella strains with visible agglutination reaction. To further verify this agglutination activity, a total of 52 flagellated Salmonella strains (23 serovars), 8 non-flagellate Salmonella strains (2 serovars) and 16 other non- Salmonella bacteria strains were used to evaluate the specificity of the MAb by direct Slide Agglutination Test (SAT). Results showed that MAb 3E2 reacted with all Salmonella strains possessing flagellum and had no cross-reaction with non-flagellate Salmonella strains or other non- Salmonella bacteria strains. Sequentially, the ability to detect the presence of Salmonella in raw samples of the MAb 3E2-based SAT method was evaluated. The conventional culture-based detection method was performed as the standard reference method for detection of Salmonella . Altogether, 369 samples collected from laying hens were tested, and the results indicated that the MAb 3E2-based SAT method could specifically detect Salmonella . Furthermore, the SAT results were obtained more quickly, as compared with the standard method. As a whole, the MAbs against the tailored conserved region of Salmonella flagellin were prepared in this study, and MAb 3E2-based SAT is a promising candidate for the flagellated Salmonella spp. rapid detection. Electronic supplement...

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“…In recent years, with the continuous improvement in the intensification of livestock husbandry and aquaculture, the density of housed animals has increased. In addition, the spread of animal infectious diseases is accelerating, and various animal epidemics have emerged ( Wu et al, 2012 ; Mellata, 2013 ; Mughini-Gras et al, 2014 ; Threlfall et al, 2014 ; Barbosa et al, 2017 ; Poulsen et al, 2017 ; Stromberg et al, 2017 ; Vinayananda et al, 2017 ). Thus, the disinfection of the livestock and poultry houses has become an important measure to prevent and control diseases.…”

Section: Introductionmentioning

confidence: 99%

Effect of Different Disinfectants on Bacterial Aerosol Diversity in Poultry Houses

Jiang

Tang

et al. 2018

Front. Microbiol.

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To better understand the effect of different disinfectants on the types and quantities of microorganisms in a broiler chicken house, five different types of disinfectants, including ozone, available chlorine, quaternary ammonium salt, glutaraldehyde, and mixed disinfectant, were used. The broiler house microbial communities were analyzed by high-throughput sequencing combined with air sampling. The results showed that the concentrations of airborne aerobic bacteria in the empty broiler houses after application of different disinfectants were significantly reduced compared to a house untreated with disinfectant (P < 0.05 or P < 0.01), and the number of inhalable particles of airborne aerobic bacteria sharply decreased after disinfection. Of the five disinfectants, the mixed disinfectant had the best disinfection efficacy on the total microbial communities (P < 0.05). A total of 508,143 high-quality sequences were obtained by high-throughput sequencing, which identified 1995 operational taxonomic units. In total, 42 phyla and 312 genera were identified. The structures of airborne microbial communities in the broiler houses after the different disinfectants were applied differed. In the house treated with the mixed disinfectant, the microbial communities containing opportunistic pathogens, such as Escherichia-Shigella, Bacillus, and Pseudomonas, had the lowest abundance, with a significant decrease compared to the house untreated with disinfectant. The alpha diversity index showed low diversity of the microbial communities in the house treated with mixed disinfectant. In contrast to the other four disinfectants, only a small amount of bacteria was detected in the air sample in the house treated with the mixed disinfectant; specifically, only four phyla were found (Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes). The mixed disinfectant produced a positive effect on disinfection for four phyla; however, it didn’t thoroughly eliminate them. At genus level, Bacillus, Arenimonas, and Shinella could not be detected in the house treated with the mixed disinfectant, but were detected in houses treated with other disinfectants. The high-throughput sequencing results revealed that the combination of multiple disinfectants exhibited a good disinfection efficacy and that this technique could disinfect the air of broiler houses. These results will help guide the development of a reasonable program for broiler house disinfection.

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Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken

Cited by 32 publications

References 18 publications

Characterization of Salmonella Dublin isolated from bovine and human hosts

Characterization of Salmonella Dublin isolated from bovine and human hosts

A promising detection candidate for flagellated Salmonella spp.

Effect of Different Disinfectants on Bacterial Aerosol Diversity in Poultry Houses

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