Structural studies of the lipopolysaccharide from the fish pathogen Aeromonas veronii strain Bs19, serotype O16.

Chemical analyses, mass spectrometry, and NMR spectroscopy were applied to study the structure of the lipopolysaccharide (LPS) isolated from Aeromonas veronii strain Bs19, serotype O16. ESI-MS revealed that the most abundant LPS glycoforms have tetra-acylated or hexa-acylated lipid A species, consisting of a bisphosphorylated GlcN disaccharide with an AraN residue as a non-stoichiometric substituent, and a core oligosaccharide composed of Hep5Hex3HexN1Kdo1P1. Sugar and methylation analysis together with 1D and 2D ¹H and ¹³C NMR spectroscopy were the main methods used, and revealed that the O-specific polysaccharide (OPS) of A. veronii Bs19 was built up of tetrasaccharide repeating units with the structure: →4)-α-D-Quip3NAc-(1→3)-α-L-Rhap-(1→4)-ß-D-Galp-(1→3)-α-D-GalpNAc-(1→. This composition was confirmed by mass spectrometry. The charge-deconvoluted ESI FT-ICR MS recorded for the LPS preparations identified mass peaks of SR- and R-form LPS species, that differed by Δm = 698.27 u, a value corresponding to the calculated molecular mass of one OPS repeating unit (6dHexNAc6dHexHexHexNAc-H2O). Moreover, unspecific fragmentation spectra confirmed the sequence of the sugar residues in the OPS and allowed to assume that the elucidated structure also represented the biological repeating unit.

Structural and immunochemical studies of the lipopolysaccharide from the fish pathogen, Aeromonas bestiarum strain K296, serotype O18.

Chemical analyses and mass spectrometry were used to study the structure of the lipopolysaccharide (LPS) isolated from Aeromonas bestiarum strain K296, serotype O18. ESI-MS revealed that the most abundant A. bestiarum LPS glycoforms have a hexa-acylated or tetra-acylated lipid A with conserved architecture of the backbone, consisting of a 1,4'-bisphosphorylated ß-(1→6)-linked D-GlcN disaccharide with an AraN residue as a non-stoichiometric substituent and a core oligosaccharide composed of Kdo1Hep6Hex1HexN1P1. 1D and 2D NMR spectroscopy revealed that the O-specific polysaccharide (OPS) of A. bestiarum K296 consists of a branched tetrasaccharide repeating unit containing two 6-deoxy-l-talose (6dTalp), one Manp and one GalpNAc residues; thus, it is similar to that of the OPS of A. hydrophila AH-3 (serotype O34) in both the sugar composition and the glycosylation pattern. Moreover, 3-substituted 6dTalp was 2-O-acetylated and additional O-acetyl groups were identified at O-2 and O-4 (or O-3) positions of the terminal 6dTalp. Western blots with polyclonal rabbit sera showed that serotypes O18 and O34 share some epitopes in the LPS. The very weak reaction of the anti-O34 serum with the O-deacylated LPS of A. bestiarum K296 might have been due to the different O-acetylation pattern of the terminal 6dTalp. The latter suggestion was further confirmed by NMR.

Characteristics of disease spectrum in relation to species, serogroups, and adhesion ability of motile aeromonads in fish.

An attempt was made to delineate the relationship between of Aeromonas species and/or serogroups and specific disease symptoms in common carp Cyprinus carpio L. and rainbow trout Oncorhynchus mykiss Walbaum. The adhesion of Aeromonas strains to various tissues in relation to disease spectrum was also tested. All strains of A. hydrophila caused skin ulcers as well as septicaemia in both carp and trout while the other strains were able to cause only skin ulcers or some specific internal lesions with or without septicaemia depending on which species and/or serogroup they represented. Disease symptoms depended also on fish species. It was found that adhesion intensity of Aeromonas strains tested was significantly higher to tissues, which were susceptible to infection with these strains. The results indicate that adhesion to various cells of the fish organism is principal marker to detect virulent Aeromonas strains. The findings presented in this study may be helpful in the appraisal of aeromonads disease risk and kind of the infection in particular fish farms by epizootiological studies or/and during routine fish examinations. They will also be useful to improve and facilitate diagnosis of bacterial fish disease.

Structure of the O-specific polysaccharide from the lipopolysaccharide of Aeromonas sobria strain Pt312.

The O-specific polysaccharide (OPS) obtained by mild-acid degradation of the lipopolysaccharide from Aeromonas sobria strain Pt312 was studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, NOESY, 1H-detected 1H,13C HSQC, and HMBC experiments. The sequence of the sugar residues was determined using 1H,1H NOESY and 1H,13C HMBC experiments. It was found that the OPS was built up of disaccharide repeating units composed of GlcpNAc and non-stoichiometrically O-acetylated Rhap residues, and had the structure.

The structure of the O-specific polysaccharide from the lipopolysaccharide of Aeromonas bestiarum strain 207.

The O-specific polysaccharide obtained by mild-acid degradation of Aeromonas bestiarum 207 lipopolysaccharide was studied by sugar and methylation analyses along with (1)H and (13)C NMR spectroscopy. The sequence of the sugar residues was determined by ROESY and HMBC experiments. It is concluded that the O-polysaccharide is composed of branched pentasaccharide repeating units of the following structure: [structure: see the text]