Practical procedure for demonstrating bacterial flagella

Kodaka, Hidemasa; Armfield, A Y; Lombard, G L; Dowell, V. R.

doi:10.1128/jcm.16.5.948-952.1982

Cited by 150 publications

(51 citation statements)

References 7 publications

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“…Spore stain was performed by Schaeffer and Fulton's method (3). Flagella stain was performed by the method described by Kodaka et al (8). Motility was examined by the hanging drop method (12).…”

Section: Methodsmentioning

confidence: 99%

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Woo

Teng

Leung

et al. 2005

Microbiology and Immunology

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Since the discovery of polymerase chain reaction (PCR) and DNA sequencing, comparison of the gene sequences of bacterial species has shown that the 16S rDNA is highly conserved within a species and among species of the same genus, and hence can be used as the new standard for classification of bacteria. Using this new standard, phylogenetic trees, based on base differences between species, are constructed; and bacteria are classified and re-classified into new genera. Recently, the use of this technique for the identification of bacterial strains with ambiguous biochemical profiles and the discovery of novel genera and species have been reported by us and other researchers (5-7, 10, 16, 18-30).Based on the results of 16S rDNA sequencing, major changes in the taxonomy of anaerobic Gram-negative bacteria have been made in recent years (2,4,9,11,15). At the moment, about 50 genera of anaerobic Gram-negative bacteria have been described, three of which were sporulated. In this study, we report the isolation of an anaerobic Gram-negative bacterial strain from the blood culture of an intravenous drug abuser. The strain, named HKU15 T , exhibited phenotypic characteristics that do not fit into patterns of any known genus and species. 16S rDNA sequencing showed that there was only about 90% base identity between the 16S rDNA of HKU15T and that of the most closely related species, Dendrosporobacter quercicolus (15). On the basis of these results we propose a new genus and species, Anaerospora hongkongensis gen. nov. sp. Abstract: A bacterium was isolated from the blood culture of an intravenous drug abuser with pseudobacteremia. The cells were strictly anaerobic, straight or slightly curved, sporulating, Gram-negative rods. It grew on sheep blood agar as non-hemolytic, pinpoint colonies after 48 hr of incubation at 37 C in an anaerobic environment. It was motile but did not produce catalase or cytochrome oxidase. 16S ribosomal DNA (rDNA) sequencing revealed three different copies of 16S rDNA sequences. More than 90% of the differences among them were due to differences in the lengths of the sequences. Phylogenetically, the bacterium is clustered with Dendrosporobacter, Sporomusa, and Propionispora, the other three genera of anaerobic, sporulating, Gram-negative rods. There were 8.6-11.1% differences between the 16S rDNA sequences of the bacterium and that of D. quercicolus, 4.7-15.1% differences between the 16S rDNA sequences of it and those of S. acidovorans, S. aerivorans, S. malonica, S. ovata, S. paucivorans, S. silvacetica, S. spaeroides, and S. termitida, and 7.6-13.1% differences between the 16S rDNA sequences of it and those of P. hippei and P. vibrioides. The G؉C content of the bacterium (mean؎SD) was 46.8؎3.2%. For these reasons, a new genus and species, Anaerospora hongkongensis gen. nov. sp. nov., is proposed, for which HKU15 T is the type strain. Anaerospora hongkongensis

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Section: Methodsmentioning

confidence: 99%

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Woo

Teng

Leung

et al. 2005

Microbiology and Immunology

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show abstract

“…All strains were initially screened by Gram stain, cytochrome oxidase test (Gerhardt et al 1981), and identified with the API 20NE Test System for non-fermenters or the API 20E for fermentative bacteria (API System, S.A., La Balme les Grottes, France) according to manufacturers' instructions. Additional tests included flagella staining (Kodaka et al 1982), pigment production on Pseudo P and Pseudo F media (Difco) after incubation at 22°C for 10 d, growth in nutrient broth at 41.5"C for 24 h, lecithinase production, and growth in 6.5% NaCl (Gilardi 197 (ATCC 7965).…”

Section: E N U M E R a T I O N Isolation A N D I D E N T I F I C A mentioning

confidence: 99%

Heterotrophic plate counts and the isolation of bacteria from mineral waters on selective and enrichment media

Manaia

Nunes

Morais

et al. 1990

Journal of Applied Bacteriology

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“…We found both LF82 and LF82 Δ LF82_314 are more enriched in capillaries containing glucose than without, and no statistically significant difference was observed, suggesting chemotaxis is intact in LF82 Δ LF82_314 . We also assessed whether the number per cell or morphology of flagella was affected by deletion of LF82_314 , using Kodaka staining [65] and transmission electron microscopy (TEM). Kodaka staining (Fig.…”

Section: Resultsmentioning

confidence: 99%

A conserved protein, BcmA, mediates motility, biofilm formation, and host colonisation in Adherent InvasiveEscherichia coli

Cogger-Ward

Collins

McLean

et al. 2019

Preprint

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25Adherent Invasive Escherichia coli (AIEC) is a non-diarrhoeagenic intestinal E. coli pathotype 26 associated with Crohn's Disease. AIEC pathogenesis is characterised by biofilm formation, 27 adhesion to and invasion of intestinal epithelial cells, and intracellular replication within 28 epithelial cells and macrophages. Here, we identify and characterise a protein in the 29 prototypical AIEC strain LF82 which is required for efficient biofilm formation and dispersal -30 LF82_p314. LF82 ΔLF82_314 have defective swimming and swarming motility, indicating 31 LF82_p314 is important for flagellar-mediated motility, and thus surface colonisation and 32 biofilm dispersal. Flagellar morphology and chemotaxis in liquid appear unaffected by 33 deletion of LF82_314, suggesting LF82_p314 does not elicit an effect on flagella biogenesis or 34 environmental sensing. Flagellar motility has been implicated in AIEC virulence, therefore we 35 assessed the role of LF82_p314 in host colonisation using a Caenorhabditis elegans model. 36 We found that LF82 ΔLF82_314 have an impaired ability to colonise the C. elegans compared 37 to wild-type LF82. Phylogenetic analysis showed that LF82_314 is conserved in several major 38 enterobacterial pathogens, and suggests the gene may have been acquired horizontally in 39 several genera. Our data suggests LF82_p314 may be a novel component in the flagellar 40 motility pathway and is a novel determinant of AIEC colonisation. Our findings have potential 41 implications not only for the pathogenesis of Crohn's Disease, but also for the course of 42 infection in several major bacterial pathogens. We propose a new designation for LF82_314, 43 biofilm coupled to motility A, or bcmA. 44 Author summary 45 Adherent Invasive Escherichia coli (AIEC) are a group of bacteria implicated in the 46 pathogenesis of Crohn's Disease, a chronic inflammatory bowel disease with no cure. Critical 47 48 colonise the host surface using specialised, propeller-like appendages called flagella. In this 49 paper, we describe a novel protein -LF82_p314 (BcmA) -which is required for efficient 50 flagella-mediated motility and surface colonisation in AIEC. Using a nematode worm 51 (Caenorhabditis elegans) infection model, we show that LF82_p314 enables effective 52 colonisation of the C. elegans gut, suggesting a role for the protein during human infection. 53 These findings indicate BcmA is significant for initial colonisation of the human gut by AIEC, 54 and therefore the onset of Crohn's Disease. 55 Introduction 56 Crohn's Disease (CD) is a chronic and relapsing inflammatory bowel disease presenting with 57 frequent bloody diarrhoea, bowel obstruction, abdominal pain, and extraintestinal 58 manifestations affecting the eyes, skin, joints, and liver (reviewed in [1-3]). CD is a complex 59 syndrome which is understood as an unchecked and inappropriate inflammatory response to 60 intestinal bacteria, potentiated by carriage of one or several of over 180 predisposing 61 immune-related alleles [4-13] and their inter...

show abstract

Practical procedure for demonstrating bacterial flagella

Cited by 150 publications

References 7 publications

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Heterotrophic plate counts and the isolation of bacteria from mineral waters on selective and enrichment media

A conserved protein, BcmA, mediates motility, biofilm formation, and host colonisation in Adherent InvasiveEscherichia coli

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