Host-driven subspeciation in the hedgehog fungus, Trichophyton erinacei, an emerging cause of human dermatophytosis.

Trichophyton erinacei is a main cause of dermatophytosis in hedgehogs and is increasingly reported from human infections worldwide. This pathogen was originally described in the European hedgehog (Erinaceus europaeus) but is also frequently found in the African four-toed hedgehog (Atelerix albiventris), a popular pet animal worldwide. Little is known about the taxonomy and population genetics of this pathogen despite its increasing importance in clinical practice. Notably, whether there are different populations or even cryptic species associated with different hosts or geographic regions is not known. To answer these questions, we collected 161 isolates, performed phylogenetic and population-genetic analyses, determined mating-type, and characterised morphology and physiology. Multigene phylogeny and microsatellite analysis supported T. erinacei as a monophyletic species, in contrast to highly incongruent single-gene phylogenies. Two main subpopulations, one specific mainly to Atelerix and second to Erinaceus hosts, were identified inside T. erinacei, and slight differences in the size of microconidia and antifungal susceptibilities were observed among them. Although the process of speciation into two lineages is ongoing in T. erinacei, there is still gene flow between these populations. Thus, we present T. erinacei as a single species, with notable intraspecies variability in genotype and phenotype. The data from wild hedgehogs indicated that sexual reproduction in T. erinacei and de novo infection of hedgehogs from soil are probably rare events and that clonal horizontal spread strongly dominates. The molecular typing approach used in this study represents a suitable tool for further epidemiological surveillance of this emerging pathogen in both animals and humans. The results of this study also highlighted the need to use a multigene phylogeny ideally in combination with other independent molecular markers to understand the species boundaries of dermatophytes. Citation: Cmoková A, Kolarík M, Guillot J, et al. 2022. Host-driven subspeciation in the hedgehog fungus, Trichophyton erinacei, an emerging cause of human dermatophytosis. Persoonia 48: 203-218. https://doi.org/10.3767/persoonia.2022.48.06.

Diagnostic value of serum galactomannan, (1,3)-ß-d-glucan, and Aspergillus fumigatus-specific IgA and IgG assays for invasive pulmonary aspergillosis in non-neutropenic patients.

Detection of serum galactomannan (GM) and (1,3)-ß-d-glucan (BG) is considered useful for non-culture diagnosis of invasive pulmonary aspergillosis (IPA) in neutropenic patients. Only few studies evaluated these seromarkers in non-neutropenic patients suspected of having IPA. The aim of this study was to evaluate both tests together with the Aspergillus fumigatus-specific serum IgG and IgA (IgAG) test for serological IPA diagnosis in non-neutropenic patients. Sera from 87 patients suspected of having IPA were retrospectively analysed. Patients were categorised into groups of proven IPA (n = 10), putative IPA (n = 31) and non-IPA colonisation (n = 46). When the GM, BG and IgAG assays were used for patients included in the study, the sensitivity/specificity/positive predictive value (PPV)/negative predictive value (NPV) were 48.8%/91.3%/83.3%/66.7%, 82.9%/73.9%/73.9%/82.9% and 75.6%/95.7%/93.9%/81.5%, respectively. Thus, the highest specificity and PPV were confirmed for the IgAG assay. Improvements in the sensitivity and NPV were achieved by "at least one positive" analysis with the GM and BG assays, with the sensitivity/specificity/PPV/NPV values being 85.0%/69.6%/71.4%/84.2%. Nevertheless, the highest sensitivity and NPV were achieved by the "at least one positive" analysis combining the GM, BG and IgAG tests (97.6% and 96.8%, respectively). The involvement of the IgAG assay could improve IPA diagnosis in non-neutropenic patients by increasing the sensitivity and NPV when combined with the GM or BG assays. Furthermore, improvement was achieved by combining the GM, BG and IgAG assays using the "at least one positive test" strategy, especially if doubt exists.

Nosocomial candidemia in the Czech Republic in 2012-2015: results of a microbiological multicentre study.

  Background: Candidemia is a severe and often life-threatening infection frequently occurring in critically ill patients. During the last decade, new therapeutic and prophylactic strategies influenced (at least in some patient subgroups) the epidemiological situation and the spectrum of causative Candida strains. The present multicentre study aimed to assess the current epidemiological situation of Candida strains causing invasive candidiasis (IC) in patients of tertiary care hospitals in the Czech Republic. MATERIAL AND METHODS: Clinical and microbiological data on patients with bloodstream yeast isolates collected in 11 tertiary care hospitals in the Czech Republic between 2012 and 2015 were analysed. The incidence of cases and species distribution were assessed by study year, age, and specialty. Moreover, an association with the Candida colonization and presence of toxigenic strains of Clostridium difficile in stool prior to blood culture positivity was analysed. For some of the strains, minimum inhibitory concentrations (MICs) of systemic antifungals were determined using standard methods. RESULTS: A total of 886 episodes of candidemia (921 yeast strains) were identified during the study period. The overall incidence per 1000 admissions was 0.40 (range 0.21-1.22 depending on the hospital). Almost half of the isolates belonged to the species Candida albicans (49.7 %), followed by Candida glabrata (15.3 %) and Candida parapsilosis (11.2 %). Non-albicans species of Candida significantly predominated in oncology wards (71.6 %) as compared to surgery (40.4 %) or internal medicine (52.0 %) departments. More than 70.0 % of patients stayed in intensive care units at the time of positive culture; in 65.0 % of patients, colonization with the same yeast species preceded blood culture positivity. In only 5.1 % of all patients, the previous presence of toxigenic strains of Clostridium difficile in stool was found. Fifty-six of the 921 yeast strains were tested for antifungal susceptibility, with an increase in MICs to azoles being observed for C. glabrata. CONCLUSION: The incidence of candidemia in the Czech Republic did not vary significantly between 2012 and 2015, and C. glabrata was the second most common yeast species after C. albicans isolated from blood.

Construction of the optical part of a time-of-flight detector prototype for the AFP detector.

We present the construction of the optical part of the ToF (time-of-flight) subdetector prototype for the AFP (ATLAS Forward Proton) detector. The ToF detector in conjunction with a 3D silicon pixel tracker will tag and measure protons originating in central exclusive interactions p + p → p + X + p, where the two outgoing protons are scattered in the very forward directions. The ToF is required to reduce so-called pileup backgrounds that arise from multiple proton interactions in the same bunch crossing at high luminosity. The background can fake the signal of interest, and the extra rejection from the ToF allows the proton tagger to operate at the high luminosity required for measurement of the processes. The prototype detector uses fused silica bars emitting Cherenkov radiation as a relativistic particle passes through it. The emitted Cherenkov photons are detected by a micro-channel plate multi-anode Photomultiplier Tube (MCP-PMT) and processed by fast electronics.

High frequency of Candida fabianii among clinical isolates biochemically identified as Candida pelliculosa and Candida utilis.

Clinical yeast isolates belonging to Candida pelliculosa, Candida utilis and Candida fabianii are difficult to distinguish in a routine mycology laboratory using common biochemical tests. The aims of this study were to determine the prevalence of C. pelliculosa, C. utilis and C. fabianii in clinical samples and to compare their minimum inhibitory concentrations (MICs) to systemic antifungals. Two hundred and forty-eight clinical yeast isolates obtained from eight large hospitals in the Czech Republic were included in this study. Identification was performed biochemically using ID 32C kit and by MALDI-TOF MS. MICs were determined using colorimetric broth dilution Sensititre YeastOne panels. From a total number of 248 isolates, 175 were identified as C. pelliculosa and 73 as C. utilis using the biochemical kit. In contrast, MALDI-TOF MS identified 222 isolates as C. fabianii, 20 as C. pelliculosa and 6 as C. utilis. The highest mean MICs were found in C. fabianii and, regardless of the studied species, in isolates from blood cultures and central venous catheters. MALDI-TOF MS revealed C. fabianii to be most prevalent in clinical samples as compared with the other studied species. Higher MIC values in C. fabianii support the importance of correct identification of this species.

[The prevalence of Candida pelliculosa, Candida utilis, and Candida fabianii in the Olomouc University Hospital: epidemiological study]. / Prevalence Candida pelliculosa, Candida utilis a Candida fabianii ve Fakultní nemocnici Olomouc - epidemiologická studie.

BACKGROUND: The aims of this study were to determine the prevalence of C. pelliculosa, C. utilis, and C. fabianii in clinical samples collected from patients hospitalized in the Olomouc University Hospital and compare their minimum inhibitory concentrations (MICs ) to nine systemic antifungals with respect to yeast species, patient age, gender, and site of infection. MATERIAL AND METHODS: Identification was performed biochemically and using mass spectrometry (MALDI-TOF MS). MICs were determined by the broth dilution method. RESULTS: Of a total of 163 clinical isolates, 119 were biochemically identified as C. pelliculosa and 44 as C. utilis. Using MALDI-TOF MS, 152 isolates were identified as C. fabianii, six as C. pelliculosa, three as C. utilis, and one as Ogataea polymorpha. The highest mean MICs were found in C. fabianii and in yeasts isolated from blood cultures and central venous catheters. CONCLUSIONS: The MALDI-TOF MS found C. fabianii to be most prevalent in clinical samples as compared with the other studied species. The probable cause of discordant results between the two methods was the absence of C. fabianii in the database of the biochemical test kit which led to misidentification of this species. Higher MIC values in C. fabianii demonstrate the importance of the precise identification of this species.

Design of Cherenkov bars for the optical part of the time-of-flight detector in Geant4.

We present the results of studies devoted to the development and optimization of the optical part of a high precision time-of-flight (TOF) detector for the Large Hadron Collider (LHC). This work was motivated by a proposal to use such a detector in conjunction with a silicon detector to tag and measure protons from interactions of the type p + p → p + X + p, where the two outgoing protons are scattered in the very forward directions. The fast timing detector uses fused silica (quartz) bars that emit Cherenkov radiation as a relativistic particle passes through and the emitted Cherenkov photons are detected by, for instance, a micro-channel plate multi-anode Photomultiplier Tube (MCP-PMT). Several possible designs are implemented in Geant4 and studied for timing optimization as a function of the arrival time, and the number of Cherenkov photons reaching the photo-sensor.

European Confederation of Medical Mycology (ECMM) epidemiological survey on invasive infections due to Fusarium species in Europe.

In order to better understand the epidemiology of fusariosis in Europe, a survey collecting information on the clinical characteristics of the patients infected by Fusarium as well as on the infecting isolates was launched. A total of 76 cases of invasive fusariosis occurring from January 2007 to June 2012 were collected and Fusarium isolates were identified by sequencing the translation elongation factor 1α (TEF) gene. Also, antifungal susceptibility was tested by broth microdilution according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Etest. Disseminated disease was considered proven in 46 cases and probable in 17 cases. Localised infection was seen in 13 cases. Gibberella fujikuroi species complex (SC), including Fusarium verticillioides and F. proliferatum, and F. solani SC were the most frequent aetiology of disseminated and localised infections, respectively. The crude mortality rate was 46 %, the highest associated with F. solani SC (67 %) and F. proliferatum (62.5 %). A wide range of antifungal susceptibilities was observed. Amphotericin B was the most potent antifungal in vitro, and itraconazole the least effective. The azoles exhibited lower minimum inhibitory concentrations (MICs) against F. verticillioides strains, with posaconazole having a slightly better performance, while F. solani SC isolates were resistant to all three azoles tested. The essential agreement between the Etest and the EUCAST method was 100 % for itraconazole and voriconazole, and 96 % for amphotericin B and posaconazole. In conclusion, we confirm that fusariosis is a rare but severe event in Europe, that G. fujikuroi SC is the predominant cause of deep infections and that different species have different antifungal in vitro susceptibility patterns.

A prospective international Aspergillus terreus survey: an EFISG, ISHAM and ECMM joint study.

OBJECTIVES: A prospective international multicentre surveillance study was conducted to investigate the prevalence and amphotericin B susceptibility of Aspergillus terreus species complex infections. METHODS: A total of 370 cases from 21 countries were evaluated. RESULTS: The overall prevalence of A. terreus species complex among the investigated patients with mould-positive cultures was 5.2% (370/7116). Amphotericin B MICs ranged from 0.125 to 32 mg/L, (median 8 mg/L). CONCLUSIONS: Aspergillus terreus species complex infections cause a wide spectrum of aspergillosis and the majority of cryptic species display high amphotericin B MICs.

Primer R108 performs best in the RAPD strain typing of three Aspergillus species frequently isolated from patients.

We evaluated the suitability of various primers for the RAPD (random amplified polymorphic DNA) accurate species identification and strain typing of Aspergillus clinical isolates. Five primers described previously were tested for their discriminatory power in three Aspergillus species (A. fumigatus, A. niger agg. and A. flavus - 23 clinical isolates and 2 reference strains). Clustering of RAPD fingerprints corresponded well with the identification based on morphological features. All isolates were resolved as different strains using the primer R108 and the RAPD protocol optimized for a Robocycler thermal cycler. RAPD with the primer R108 thus can be considered to be a valuable, simple and powerful tool for identification and strain delineation of Aspergillus spp.

Efficacy of Preemptive Gabapentin for Lower Extremity Orthopedic surgery under Subarachnoid Block.

INTRODUCTION: Gabapentin is one of the adjuncts when given before surgery as a multi modal analgesia regimen has shown to decrease pain scores and opioid consumption in the first 24 hour after surgery. The aim of the study is to assess total opioid consumption in first 24 hour postoperatively after giving 600mg of preemptive oral Gabapentin in lower extremity orthopedic surgery done under subarachnoid block. METHODS: A randomized single blinded study was conducted for duration of 6 months after approval from institutional review board. A total of 52 American Society of Anesthesiology physical status grade I and II cases were included in the study of which 26 patients received oral Gabapentin two hours prior to surgery and 26 patients in the control group. They were evaluated postoperatively for total opioid consumption in first 24 hours, Visual Analogue Score after 2, 4, 6 and 24 hours at rest. RESULTS: Total opioid consumption in first 24 hours was 74.13 ± 27.78 mg in Gabapentin group versus 123.53 ± 64.48 mg in Control (p = 0.001). VAS score was 1.23 ± 1.47 in Gabapentin group versus 2.12 ± 1.58 in Control group (p=0.04) at 2 hours and 2.19 ± 0.40 in Gabapentin group versus 2.77 ± 1.17 in control group (p=0.02) at 24 hours postoperatively at rest which were found to be statistically significant. Incidence of sedation was minimal and comparable in both groups. CONCLUSIONS: Oral Gabapentin 600mg given two hours before surgery reduces total opioid consumption in first 24 hours after surgery and also reduces Visual Analogue score for pain postoperatively at rest in 2 and 24 hours with minimum sedation.

Invasive Candida infections in surgical patients in intensive care units: a prospective, multicentre survey initiated by the European Confederation of Medical Mycology (ECMM) (2006-2008).

A prospective, observational, multicentre study of invasive candidosis (IC) in surgical patients in intensive care units (ICUs) was conducted from 2006 to 2008 in 72 ICUs in 14 European countries. A total of 779 patients (62.5% males, median age 63 years) with IC were included. The median rate of candidaemia was 9 per 1000 admissions. In 10.8% the infection was already present at the time of admission to ICU. Candida albicans accounted for 54% of the isolates, followed by Candida parapsilosis 18.5%, Candida glabrata 13.8%, Candida tropicalis 6%, Candida krusei 2.5%, and other species 5.3%. Infections due to C. krusei (57.9%) and C. glabrata (43.6%) had the highest crude mortality rate. The most common preceding surgery was abdominal (51.5%), followed by thoracic (20%) and neurosurgery (8.2%). Candida glabrata was more often isolated after abdominal surgery in patients ≥60 years, and C. parapsilosis was more often isolated in neurosurgery and multiple trauma patients as well as children ≤1 year of age. The most common first-line treatment was fluconazole (60%), followed by caspofungin (18.7%), liposomal amphotericin B (13%), voriconazole (4.8%) and other drugs (3.5%). Mortality in surgical patients with IC in ICU was 38.8%. Multivariate analysis showed that factors independently associated with mortality were: patient age ≥60 years (hazard ratio (HR) 1.9, p 0.001), central venous catheter (HR 1.8, p 0.05), corticosteroids (HR 1.5, p 0.03), not receiving systemic antifungal treatment for IC (HR 2.8, p <0.0001), and not removing intravascular lines (HR 1.6, p 0.02).

Rate of transmission and endogenous origin of Candida albicans and Candida glabrata on adult intensive care units studied by pulsed field gel electrophoresis.

We determined the relative roles of endogenous origin and patient-to-patient transmission in Candida colonization of patients on adult intensive care units (ICU). A total of 48 Candida albicans and 18 Candida glabrata strains from various clinical samples of 28 long-term patients, hospitalized in two neurological ICUs between April and June 1999, were typed using pulsed field gel electrophoresis (PFGE). Three patients were co-colonized by both C. albicans and C. glabrata strains. Twenty-four C. albicans and 17 C. glabrata karyotypes were defined. The colonization was found to be polyclonal in six C. albicans and five C. glabrata patients. Twenty-six patients (93%) carried strains, which were not detected in other patients hospitalized at the same time, i.e. they were colonized by unique C. albicans and C. glabrata strains. Only two patients, who were hospitalized during the same period of time, although in different rooms of the same ICU, shared strains with an identical PFGE type, indicating possible patient-to-patient transmission. Patient-to-patient transmission of yeasts played a minor role on these ICUs.

Influence of an extended incubation period on values of minimum inhibitory concentrations of antibiotics in Stenotrophomonas maltophilia clinical strains.

In 106 Stenotrophomonas maltophilia clinical strains the susceptibility to 19 kinds of antibiotics was tested by the broth dilution micromethod at 24 h and 48 h incubation. Isolated strains demonstrated the lowest frequency of resistance to cotrimoxazole (7.5% of resistant strains at 24 h incubation and 18.9% at 48 h), ofloxacin (13.2% and 30.2%), ciprofloxacin (19.8% and 50.9%) and to cefoperazone/sulbactam (20.8% and 37.7%). The smallest growth of the number of resistant strains after extended incubation was recorded in gentamicin (by 10.4%), ceftazidime (by 11.3%) and cotrimoxazole (by 11.4%). On the contrary, the largest growth of resistance was demonstrated in cefoperazone and ciprofloxacin (by 31.1%). Average values of the growth of minimum inhibitory concentrations (MICs) were lowest in ciprofloxacin and ofloxacin (2.3 times) and highest in piperacillin/tazobactam (4.5 times) and piperacillin (5.0 times). As far as the stability of MIC is concerned, the largest occurrence of strains with the MIC growth doubled as a maximum was found in ceftazidime (78.4%), ofloxacin (76.1%) and ciprofloxacin (75.3%), the smallest in piperacillin/tazobactam (43.2%) and piperacillin (38.9%). The importance of incubation extended to 48 h during the testing of S. maltophilia strains was noted for correctly setting their susceptibility to antibiotics.

Occurrence of variants with temperature-dependent susceptibility (TDS) to antibiotics among Stenotrophomonas maltophilia clinical strains.

Susceptibility to 20 antibiotics was tested in 104 Stenotrophomonas maltophilia strains at 37 and 30 degrees C by means of a dilution micromethod to verify the phenomenon of temperature-dependent susceptibility (TDS). Trimethoprim-sulfamethoxazole, pefloxacin and ofloxacin were the most active preparations at 37 degrees C (93, 90, and 86% of susceptible strains, respectively), whilst trimethoprim-sulfamethoxazole, cefoperazone-sulbactam and pefloxacin performed best at 30 degrees C (94, 94, and 76% of susceptible strains, respectively). Variants 37TDS (minimum inhibitory concentration, MIC, of tested antibiotics at least 4-times lower at 37 than at 30 degrees C) occurred in 60%. Variants 30TDS (at least 4-times lower value of MIC at 30 than at 37 degrees C) were found in 7.7%. Both variants in susceptibility to tested antibiotics appeared in 23.1%, whilst neither of them was observed in 9.6%. The 37TDS phenomenon was recorded most of all with gentamicin (51% of strains), amikacin (47), colistin (44) and tetracycline (34). The 30TDS phenomenon was found particularly with cefoperazone-sulbactam (16.0% of strains) and colistin (10.0%). The above phenomena may be due to changes in membrane permeability, temperature-dependent ribosomal changes, and insufficient adaptation to higher temperatures of some strains of the originally environmental species S. maltophilia.

Secreted aspartate proteinases, a virulence factor of Candida spp.: occurrence among clinical isolates.

Production of secreted aspartate proteinases was determined in a set of 646 isolates of Candida and non-Candida yeast species collected from 465 patients of the University Hospital in Olomouc (Czechia) in the period 1995-2002, and Candida samples obtained from 64 healthy volunteers using solid media developed for this purpose. Using random amplified polymorphic DNA analysis (RAPD) 79 Candida isolates from blood were analyzed to show potential relationships between clustering of the fingerprints and extracellular proteolytic activity of these strains. C. albicans, C. tropicalis and C. parapsilosis possess always proteolytic activity while non-Candida species did not display any proteolysis. A tight relationship between fingerprints and extracellular proteolysis in the Candida isolates was not shown. A remarkable consistency between fingerprint clusters and proteolysis occurred in a subset of C. parapsilosis samples. Suboptimal pH of the growth medium was shown to facilitate the investigation of potential co-incidence of genotypic and phenotypic traits.

[Personal experience with classification of yeast microorganisms. I. The combined biotyping method of Mencl and Otcenásek and typing using the "killer" phenomenon]. / Nase zkusenosti s klasifikací kvasinkovitých mikroorganismu. I. Kombinovaná biotypizacní metoda podle Mencla a Otcenáska a typizace s vyuzitím "killer" fenoménu.

Using a group of 150 isolates of Candida albicans, C. glabrata, C. krusei, C. tropicalis, C. parapsilosis a C. kefyr the differentiating capacity of two biotyping systems was tested-the combined method according to Mencl and Otcenásek, and typing using the so-called killer phenomenon. With the combined method comparable results with the original work of the authors were obtained. This applies to the number of biotypes as well as to the ratio of the dominant biotype. As regards the differentiating characteristics of different biotypes the two studies differed fundamentally. As to typing, using the "killer" phenomenon, its practical usefulness was tested, the differentiating capacity of the method was, however, very much influenced by the small number of available killer-positive yeast strains.

Analysis of Health Sector Budget of Nepal.

INTRODUCTION: Primarily, health sector connects two segments - medicine and public health, where medicine deals with individual patients and public health with the population health. Budget enables both the disciplines to function effectively. The Interim Constitution of Nepal, 2007 has adapted the inspiration of federalism and declared the provision of basic health care services free of cost as a fundamental right, which needs strengthening under foreseen federalism. METHODS: An observational retrospective cohort study, aiming at examining the health sector budget allocation and outcome, was done. Authors gathered health budget figures (2001 to 2013) and facts published from authentic sources. Googling was done for further information. The keywords for search used were: fiscal federalism, health care, public health, health budget, health financing, external development partner, bilateral and multilateral partners and healthcare accessibility. The search was limited to English and Nepali-language report, articles and news published. RESULTS: Budget required to meet the population's need is still limited in Nepal. The health sector budget could not achieve even gainful results due to mismatch in policy and policy implementation despite of political commitment. CONCLUSIONS: Since Nepal is transforming towards federalism, an increased complexity under federated system is foreseeable, particularly in the face of changed political scenario and its players. It should have clear goals, financing policy and strict implementation plans for budget execution, task performance and achieving results as per planning. Additionally, collection of revenue, risk pooling and purchasing of services should be better integrated between central government and federated states to horn effectiveness and efficiency.

Health system through the eyes of a doctor from rural Nepal.

Its almost 30 years of declaration of Alma-Ata for primary healthcare policy the health system in Nepal still facing shortage of trained medical doctors and health professionals reaching remote and rural part of the country to provide quality health services. There are number of issues such as financial or non-financial incentives, professional advancements, educational opportunities and workplace environment. Healthcare delivery system in Nepal is failing to meet the healthcare need of the general public and needs discussion and revision. However, despite of so many challenges more doctors are willing to work in the remote and rural Nepal. The government has to come out with effective planning and policy regarding health system and human resource for health. In this context, an attempt has been made for a analytical perspective from a medical doctor point of view to highlight some of the pertinent local and policy related issues to improve Health System in Nepal.