First analysis by <scp>MALDI</scp>‐<scp>TOF MS</scp> technique of <i>Chryseobacterium</i> species relevant to aquaculture

Vela, Ana Isabel; Kostrzewa, Markus; Zamora, Leydis; Casamayor, A.; Domı́nguez, Lucas; Fernández‐Garayzábal, J. F.

doi:10.1111/jfd.12706

Cited by 7 publications

(5 citation statements)

References 29 publications

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“…in aquaculture. 22 Consequently, the consistency of MALDI-TOF MS identification results in our study was evaluated considering the 2 best scores provided by MALDI-TOF MS Biotyper. Following this criterion, all isolates of S. aureus subsp.…”

Section: Discussionmentioning

confidence: 99%

“…The MALDI Biotyper software generated a score value–based ranking list of the 10 best identification matches. For interpretation of the identification results, 2 variables were considered 22 : 1) score values following the MALDI-TOF MS Biotyper Compass (⩾2.000 = identification at species level with high confidence; 1.999–1.800 = identification with low confidence; ⩽1.799 = non-reliable identification), and 2) consistency ranking list of the first 2 best matches (A = the correct subspecies is the unique subspecies with a score value ⩾2.000; B = the correct subspecies is first ranked but the other subspecies in the second rank also gave a score value ⩾2.000; C = first and second matches have score values ⩾2.000 but the correct species is second ranked; D = first and second matches have score values <2.000 [correct genus identification]).…”

Section: Methodsmentioning

confidence: 99%

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Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles

et al. 2018

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Staphylococcus aureus encompasses 2 subspecies (aureus and anaerobius) with significant differences in their epidemiology and pathogenicity. We evaluated the suitability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the rapid identification of both subspecies using a panel of 52 S. aureus isolates (30 subsp. anaerobius and 22 subsp. aureus) recovered from different origins, countries, and years. The on-board library identification system correctly identified 42 of 52 (81%) S. aureus isolates at the species level with score values >2.0. Limited performance was observed for differentiation of S. aureus subspecies (particularly subsp. anaerobius). Visual inspection of MALDI-TOF MS profiles identified 5 subspecies-specific mass peaks (m/z 3430 and 6861 in S. aureus subsp. anaerobius, and m/z 4046, 6890, and 8093 in S. aureus subsp. aureus) with 100% sensitivity and specificity values, which is potentially useful for differentiating these subspecies. The suitability of 3 models, Genetic Analysis (GA), Quick Classifier (QC), and Supervised Neural Network, for automatic identification of both subspecies was evaluated using the Recognition Capability (RC) and Cross Validation (CV) values provided by the on-board ClinProTools software. The GA and QC models reached RC and CV values of 100%. Both models were externally validated using a panel of 26 S. aureus isolates of both subspecies, with both models correctly classifying all isolates of both subspecies. MALDI-TOF MS coupled with ClinProTools software represents a rapid and simple approach for S. aureus subspecies discrimination.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles

et al. 2018

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“…They include polymerase chain reaction (PCR) and gene sequencing, which have largely replaced techniques involving the identification of microorganisms using phenotypic methods. The technique of identifying microorganisms based on their protein profile by MALDI-TOF MS is increasingly used for various bacterial isolates, including Campylobacter, Lactobacillus, and Enterococcus [18][19][20], as well as in the case of non-fermenting bacteria, including those of the family Weeksellaceae, which includes the genus Riemerella [21,22]. Mass spectrometry can also be successfully used to identify strains of R. anatipestifer, and the sensitivity of this method is reflected by the results of the present study, in which we obtained a high identification score in MALDI-TOF MS for nearly all bacterial isolates.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Riemerella anatipestifer Strains Isolated from Various Poultry Species in Poland

Nowaczek,

Dec,

Stępień-Pyśniak

et al. 2023

Antibiotics

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Riemerella anatipestifer (R. anatipestifer) is one of the common pathogens found in poultry flocks, resulting in serious economic losses for the poultry industry due to high mortality, reduced growth rate, poor feed conversion, increased condemnations, and high treatment costs. The aim of this study was to phenotypically characterize phylogenetic relationships and assess the presence of resistance gene strains of R. anatipestifer obtained from various poultry species in Poland. A total of 57 isolates of Riemerella were included in this study. A polymerase chain reaction (PCR) and matrix assisted laser desorption ionization mass spectrometry (MALDI-TOF MS) were used for identification of the strains. The phylogenetic relationship of the R. anatipestifer isolates was determined by analysing the rpoB gene sequence. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in liquid media. All of the field strains of R. anatipestifer were grouped into one of two clades resulting from rpoB gene sequencing. High MIC50 and MIC90 values were obtained for gentamycin, amikacin, and colistin. Low MIC50 and MIC90 values were obtained for amoxicillin cefuroxime, cefoperazone, piperacillin, and trimethoprim/sulfamethoxazole. Among the resistance genes, tet(X) and ermF were identified most frequently. This is the first phenotypic characterization of R. anatipestifer strains obtained from poultry flocks in Poland.

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“…were subcultured on Columbia Agar plates supplemented with 5% sheep blood (Cos, Biomerieux, Marcy-l'Etoile, France) and incubated for 48 h at 37 ºC under microaerophilic conditions. Subcultured isolates were then analysed by matrix assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry at the Microbial Identification and Characterization Unit at the Complutense University of Madrid (Spain) following the procedures detailed in Perez-Sancho et al [ 53 ].…”

Section: Methodsmentioning

confidence: 99%

Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain

Pena-Fernández,

Kortabarria,

Hurtado

et al. 2024

BMC Vet Res

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Background Bovine genital campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis (Cfv) including its biovar intermedius (Cfvi). This sexually transmitted disease induces early reproductive failure causing considerable economic losses in the cattle industry. Using a collection of well-characterized isolates (n = 13), C. fetus field isolates (n = 64) and saprophytic isolates resembling Campylobacter (n = 75) obtained from smegma samples of breeding bulls, this study evaluated the concordance of the most used phenotypic (H2S production in cysteine medium and 1% glycine tolerance) and molecular (PCR) methods for the diagnosis of BGC and assessed possible cross-reactions in the molecular diagnostic methods. Results Characterization at the subspecies level (fetus vs. venerealis) of C. fetus isolated from bull preputial samples using phenotypic and molecular (PCR targeting nahE and ISCfe1) methods showed moderate concordance (κ = 0.462; CI: 0.256–0.669). No cross-reactions were observed with other saprophytic microaerophilic species or with other Campylobacter species that can be present in preputial samples. Whole genome sequencing (WGS) of discrepant isolates showed 100% agreement with PCR identification. For the differentiation of Cfv biovars, comparison of the H2S test (at 72 h and 5 days of incubation) and a PCR targeting the L-cysteine transporter genes showed higher concordance when H2S production was assessed after 5 days (72 h; κ = 0.553, 0.329–0.778 CI vs. 5 days; κ = 0.881, 0.631–1 CI), evidencing the efficacy of a longer incubation time. Conclusions This study confirmed the limitations of biochemical tests to correctly identify C. fetus subspecies and biovars. However, in the case of biovars, when extended incubation times for the H2S test (5 days) were used, phenotypic identification results were significantly improved, although PCR-based methods produced more accurate results. Perfect agreement of WGS with the PCR results and absence of cross-reactions with non-C. fetus saprophytic bacteria from the smegma demonstrated the usefulness of these methods. Nevertheless, the identification of new C. fetus subspecies-specific genes would help to improve BGC diagnosis.

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First analysis by MALDI‐TOF MS technique of Chryseobacterium species relevant to aquaculture

Cited by 7 publications

References 29 publications

Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles

Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles

Characterization of Riemerella anatipestifer Strains Isolated from Various Poultry Species in Poland

Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain

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