Comparison of MALDI-TOF mass spectrometry and rpoB gene sequencing for the identification of clinical isolates of Aeromonas spp.

Kitagawa, Daisuke; Suzuki, Yuki; Abe, Noriyuki; Ui, Koji; Suzuki, Keisuke; Yamashita, Takahiro; Sakaguchi, Atsuko; Suzuki, Soma; Masuo, Kazue; Nakano, Akiyo; Nakano, Ryuichi; Sato, Masatoshi; Maeda, Koichi; Nakamura, Fumihiko; Yano, Hisakazu

doi:10.1016/j.heliyon.2022.e11585

Cited by 9 publications

(11 citation statements)

References 29 publications

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“…infection was monomicrobial in most cases (93%). In contrast, in previous studies evaluating traveler’s or children’s diarrhea, multiple pathogens were detected in a higher percentage of cases ( 31 , 32 , 37 ), probably due to differences in the study population. Our survey is free of the bias of patient recruitment and reveals a bimodal case distribution, with a peak in children younger than 15 years and a second large peak in subjects older than 45.…”

Section: Discussioncontrasting

confidence: 78%

“…In the current study, Aeromonas spp. were identified using the rpoB gene sequencing method, a procedure considered much more accurate than biochemical identification methods or 16S rRNA gene sequencing ( 17 , 37 , 38 ). In agreement with recent studies, A. caviae was recognized as humans' predominating species associated with diarrhea ( 32 , 39 ).…”

Section: Discussionmentioning

confidence: 99%

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Prevalence and virulence potential of Aeromonas spp. isolated from human diarrheal samples in North East Italy

Bernabè,

Brun,

Pietra

et al. 2023

Microbiol Spectr

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Aeromonas spp. are emerging human pathogens causing intestinal and extra-intestinal infections. Since their relevance in Western Europe as gastrointestinal pathogens is not well established, we investigated Aeromonas spp. prevalence in diarrheal fecal samples in an Italian University Hospital and characterized the virulence mechanisms of the isolates. Aeromonas spp. isolated from diarrheic stools using standard culture methods were identified by molecular techniques. Antimicrobial resistance was assessed by the micro broth dilution. Toxins, flagella, and type III secretion system genes were evaluated by polymerase chain reaction. Biofilm was quantified by crystal-violet staining. Interaction with human intestinal epithelial cells (Caco-2) was assessed by quantifying adhesion, interleukin (IL)-8 secretion, and epithelial barrier integrity. Aeromonas spp. represented 20.6% of bacterial pathogens isolated from diarrheic feces, the second most common enteropathogen. A. cavieae constituted 75% of the identified species, showing a relatively low clustering value. About 52% of Aeromonas isolates showed resistance to amikacin, whereas only 7.5% showed multiple drug resistance. Four or more virulence genes were identified in 66.7% of A. cavieae isolates and 100% of A. dakensis. Aeromonas isolates (82.5%) showed moderate or important biofilm-producing ability. Adhesion to Caco-2 cells correlated to fla + gene, whereas ascV+ and aexU + strains significantly induced IL-8 release from Caco-2. Aeromonas aer+ strains caused ZO1 and occluding redistribution and a significative reduction in trans-epithelial resistance. Aeromonas spp. emerge as relevant human intestinal pathogens with a disparate arsenal of pathogenicity factors causing diarrhea through different mechanisms. IMPORTANCE In this work, we demonstrate the epidemiologic relevance of the Aeromonas genus as the cause of infective diarrhea in North East Italy, both in children and adult subjects, with the significative presence of highly pathogenic strains. Aeromonas strains possess a heterogeneous armamentarium of pathogenicity factors that allows the microbe to affect a wide range of human intestinal epithelial cell processes that justify the ability to induce diarrhea through different mechanisms and cause diseases of variable severity, as observed for other gastrointestinal pathogens. However, it remains to be determined whether specific genotype(s) are associated with clinical pictures of different severity to implement the diagnostic and therapeutic approaches for this relevant enteric pathogen.

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Section: Discussioncontrasting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Prevalence and virulence potential of Aeromonas spp. isolated from human diarrheal samples in North East Italy

Bernabè,

Brun,

Pietra

et al. 2023

Microbiol Spectr

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“…At present, according to the research progress in China and abroad, MALDI-TOF MS has been used in clinical detection and basic research for microbial typing, bacterial drug resistance, epidemiological analysis, and recombinant protein expression in bacteria, not only improving the efficiency and accuracy of microbial detection but also being used to study microorganisms at the proteomics level. However, MALDI-TOF-MS also has some defects, which need to be combined with other means for bacterial identification [25][26][27]. In the present study, 16S rRNA sequence analysis, physiological and biochemical determination, and MALDI-TOF-MS technology were used to identify the pathogenic bacteria ZZ051 of channel catfish, increasing the accuracy of the results.…”

Section: Discussionmentioning

confidence: 97%

Isolation, Identification, and Pathogenicity of Aeromonas veronii, the Causal Agent of Hemorrhagic Septicemia in Channel Catfish (Ictalurus punctatus) in China

Qin

et al. 2022

Fishes

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Herein, we isolated the pathogenic strain ZZ051 from hemorrhagic channel catfish (Ictalurus punctatus). Physiological and biochemical identification, 16S rRNA gene sequence analysis, and MALDI-TOF-MS showed that the ZZ051 strain was Aeromonas veronii. After artificial infection, the diseased fish showed symptoms similar to the natural disease, and the characteristics of the bacteria reisolated from the tissues were the same as those of the original infection, indicating that the isolated strain ZZ051 was the pathogen responsible for the channel catfish disease. The ZZ051 isolate was highly sensitive to enrofloxacin but resistant to florfenicol. This study provided a theoretical basis for preventing and controlling hemorrhagic disease in channel catfish.

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“…The thirty-one Aeromonas isolates, including 11 isolates of Aeromonas hydrophila, 10 isolates of A. caviae, and 10 isolates of A. sobria, were identi ed by MALDI-TOF MS (Bruker Daltonik, Bremen, Germany) and rpoB sequencing [12][13]. The species were identi ed by DNA sequence matching of rpoD and gyrB (or rpoB if necessary).…”

Section: Bacterial Isolatesmentioning

confidence: 99%

Synergistic effect of repurposed mitomycin C in combination with antibiotics against Aeromonas infection: in vitro and in vivo studies.

Yeh,

Chen,

Lai

et al. 2024

Preprint

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Background Aeromonas infections pose a significant threat, presenting a potentially life-threatening condition associated with high mortality rates. This study investigates the potential of mitomycin C (MMC), an anticancer drug, as a novel antimicrobial agent against Aeromonas infections. Methods The minimum inhibitory concentrations (MICs) of MMC and antibiotics against clinical isolates of Aeromonas species were assessed, using broth microdilution. Synergistic effects of MMC with antibiotics were assessed through time-kill studies. Additionally, we analyzed MMC's intracellular killing effects using a representative Aeromonas isolate. Efficacy of combined therapies was evaluated in neutropenic mice model. Results We found that only three antibiotics-ciprofloxacin (CIP), levofloxacin (LVX), and tigecycline (TGC) maintained susceptibility rates exceeding 90% under high inoculum conditions. Significantly, in vitro investigations demonstrated a synergistic effect over a 24-hour period when combining the 1/8x MIC of MMC with the 1x or 1/2x MICs of LVX, in a time–kill study. Furthermore, in vivo in neutropenic mice, the combination of 2 mg/kg and 1 mg/kg MMC with LVX resulted in 100% and 80% survival, respectively, whereas treatment with either drug alone resulted in 0% survival rate. Conclusions These findings suggest that MMC can be repurposed as an antimicrobial agent, particularly with a minimized dose and in combination with antibiotics.

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Comparison of MALDI-TOF mass spectrometry and rpoB gene sequencing for the identification of clinical isolates of Aeromonas spp.

Cited by 9 publications

References 29 publications

Prevalence and virulence potential of Aeromonas spp. isolated from human diarrheal samples in North East Italy

Prevalence and virulence potential of Aeromonas spp. isolated from human diarrheal samples in North East Italy

Isolation, Identification, and Pathogenicity of Aeromonas veronii, the Causal Agent of Hemorrhagic Septicemia in Channel Catfish (Ictalurus punctatus) in China

Synergistic effect of repurposed mitomycin C in combination with antibiotics against Aeromonas infection: in vitro and in vivo studies.

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