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Species Identification and In Vitro Antifungal Susceptibility of Paecilomyces/Purpureocillium Species Isolated from Clinical Respiratory Samples: A Multicenter Study

ABSTRACT: Paecilomyces spp. are emerging fungal pathogens, where Paecilomyces lilacinus and Paecilomyces variotii are the most reported species. Taxonomic and phylogenetic revisions in this genus have shown that P. variotii represents a species complex, whereas P. lilacinus is related to another genus called Purpureocillium. The aims of this study were to identify clinical isolates of Paecilomyces spp. at the species level, and to determine their antifungal susceptibility profiles. 70 clinical Paecilomyces spp. isolates were identified by MALDI-TOF Mass Spectrometry (MS) and by multilocus rDNA genes sequencing including ITS and the D1/D2 genes. Among the 70 Paecilomyces spp. isolates, 28 were identified as P. lilacinum, 26 as P. variotii stricto sensu, and 16 as P. maximus. For antifungal susceptibility testing, Minimal Inhibitory Concentrations (MICs) or Minimal Effective Concentrations (MECs) were determined for 8 antifungals. All P. lilacinum isolates had high MICs and MECs of amphotericin B and echinocandins, respectively, unlike P. variotii and P. maximus. For azole drugs, MICs were molecule- and species- dependent. The differences in in vitro susceptibility to antifungals underline the importance of accurate species identification. The MALDI–TOF MS can be a good alternative in routine laboratory to ensure fast identification of Paecilomyces spp. and P. lilacinum.

SUBMITTER: Monpierre L

PROVIDER: S-EPMC9321559 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundAspergillus species cause a variety of serious clinical conditions with increasing trend in antifungal resistance. The present study aimed at evaluating hospital epidemiology and antifungal susceptibility of all isolates recorded in our clinical database since its implementation.MethodsData on date of isolation, biological samples, patients' age and sex, clinical settings, and antifungal susceptibility tests for all Aspergillus spp. isolated from 2015 to 2022 were extracted from the clinical database. Score test for trend of odds, non-parametric Mann Kendall trend test and logistic regression analysis were used to analyze prevalence, incidence, and seasonality of Aspergillus spp. isolates.ResultsA total of 1126 Aspergillus spp. isolates were evaluated. A. fumigatus was the most prevalent (44.1%) followed by A. niger (22.3%), A. flavus (17.7%) and A. terreus (10.6%). A. niger prevalence increased over time in intensive care units (p-trend = 0.0051). Overall, 16 (1.5%) were not susceptible to one azole compound, and 108 (10.9%) to amphotericin B, with A. niger showing the highest percentage (21.9%). The risk of detecting A. fumigatus was higher in June, (OR = 2.14, 95% CI [1.16; 3.98] p = 0.016) and reduced during September (OR = 0.48, 95% CI [0.27; 0.87] p = 0.015) and October as compared to January (OR = 0.39, 95% CI [0.21; 0.70] p = 0.002. A. niger showed a reduced risk of isolation from all clinical samples in the month of June as compared to January (OR = 0.34, 95% CI [0.14; 0.79] p = 0.012). Seasonal trend for A. flavus showed a higher risk of detection in September (OR = 2.7, 95% CI [1.18; 6.18] p = 0.019), October (OR = 2.32, 95% CI [1.01; 5.35] p = 0.048) and November (OR = 2.42, 95% CI [1.01; 5.79] p = 0.047) as compared to January.ConclusionsThis is the first study to analyze, at once, data regarding prevalence, time trends, seasonality, species distribution and antifungal susceptibility profiles of all Aspergillus spp. isolates over a 8-year period in a tertiary care center. Surprisingly no increase in azole resistance was observed over time.

Project description:Antibiotic resistance in Ureaplasma urealyticum/Ureaplasma parvum and Mycoplasma hominis is an issue of increasing importance. However, data regarding the susceptibility and, more importantly, the clonality of these organisms are limited. We analyzed 140 genital samples obtained in Bern, Switzerland, in 2014. Identification and antimicrobial susceptibility tests were performed by using the Mycoplasma IST 2 kit and sequencing of 16S rRNA genes. MICs for ciprofloxacin and azithromycin were obtained in broth microdilution assays. Clonality was analyzed with PCR-based subtyping and multilocus sequence typing (MLST), whereas quinolone resistance and macrolide resistance were studied by sequencing gyrA, gyrB, parC, and parE genes, as well as 23S rRNA genes and genes encoding L4/L22 ribosomal proteins. A total of 103 samples were confirmed as positive for U. urealyticum/U. parvum, whereas 21 were positive for both U. urealyticum/U. parvum and M. hominis. According to the IST 2 kit, the rates of nonsusceptibility were highest for ciprofloxacin (19.4%) and ofloxacin (9.7%), whereas low rates were observed for clarithromycin (4.9%), erythromycin (1.9%), and azithromycin (1%). However, inconsistent results between microdilution and IST 2 kit assays were recorded. Various sequence types (STs) observed previously in China (ST1, ST2, ST4, ST9, ST22, and ST47), as well as eight novel lineages, were detected. Only some quinolone-resistant isolates had amino acid substitutions in ParC (Ser83Leu in U. parvum of serovar 6) and ParE (Val417Thr in U. parvum of serovar 1 and the novel Thr417Val substitution in U. urealyticum). Isolates with mutations in 23S rRNA or substitutions in L4/L22 were not detected. This is the first study analyzing the susceptibility of U. urealyticum/U. parvum isolates in Switzerland and the clonality outside China. Resistance rates were low compared to those in other countries. We hypothesize that some hyperepidemic STs spread worldwide via sexual intercourse. Large combined microbiological and clinical studies should address this important issue.

Project description:Background and purposeOpportunistic fungal infections have been on a growing trend since the last two decades. Among the opportunistic fungal agents, Candida species, Cryptococcus neoformans, and Aspergillus fumigatus account for most of the life-threatening infections in immunocompromised individuals. Regarding this, the present study aimed to investigate the molecular identification and antifungal susceptibility pattern of Candida species isolated from HIV-infected patients.Materials and methodsThis study was conducted on 80 clinical samples collected from HIV-infected patients with suspected candidiasis referring to Tagore Medical College and Hospital, Rathinamangalam and Government Hospital of Thoracic Medicine, in Chennai, India, for 18 months (i.e., May 2016-December 2017). Phenotypic and molecular identification was accomplished using internal transcribed spacer region 1 (ITS1) and ITS4 primers. The antifungal susceptibility pattern of the isolates against four antifungal agents was also determined by both disk diffusion and broth dilution methods.ResultsIn the present study, the prevalence of candidiasis was obtained as 75% (n=60). Candida tropicalis was the predominant identified species. All the emerging species (i.e., Kodamaea ohmeri, Hanseniaspora opuntiae, and C. orthopsilosis) were identified through molecular identification since the phenotypic identification was inconclusive. In terms of the susceptibility pattern, 63.3% and 18.3% of the isolates were resistant to fluconazole and voriconazole, respectively. Candida albicans was also found to be resistant to amphotericin B.ConclusionMolecular assay led to the identification of K. ohmeri, H. opuntiae, and C. orthopsilosis, which were multidrug-resistant. This study highlighted the need for the prompt and timely identification of clinical yeast isolates given the emergence of many rare species and their capability of causing life-threatening infections and outbreaks. In the laboratories where molecular diagnostic methods are not available, alternative services of reference laboratories can be utilized as cost-effective measures. With regard to the growing prevalence of antifungal drug resistance, antifungal susceptibility testing should be made mandatory for effective patient management.

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Species Identification and In Vitro Antifungal Susceptibility of Paecilomyces/Purpureocillium Species Isolated from Clinical Respiratory Samples: A Multicenter Study

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