Publications by authors named "Sebastian Gnat"

57 Publications

New Reference Genes for qRT-PCR Analysis as a Potential Target for Identification of in Different Culture Conditions.

Pathogens 2021 Oct 21;10(11). Epub 2021 Oct 21.

Department of Mycology and Genetics, Faculty of Biological Sciences, University of Wroclaw, 51-148 Wroclaw, Poland.

Dermatophytes are a group of filamentous fungi infecting skin, hair, and nails that raise great diagnostic difficulties. qRT-PCR is a reliable technique for quantifying gene expression with increasingly frequent use in mycological diagnostics. Knowledge of genes and molecular markers with potential to be used in the identification of dermatophytes is of great importance for the development of this branch of diagnostics. In this article, the suitability of six candidate reference genes (, , , , , and ) was investigated for gene expression analysis in the dermatophyte , which was cultured in various mycological media that are commonly used in a diagnostic laboratory, i.e., Sabouraud, potato dextrose, and keratin-supplemented MM-Cove. The different culture conditions are extremely important factors for the growth and physiology of dermatophytes. Gene expression stability was evaluated using geNorm, NormFinder, BestKeeper, and RefFinder algorithms. Regarding the stability of expression, was the most stable housekeeping gene; hence, this gene is recommended for future qRT-PCR studies on strains. These results allow us to conclude that the gene can be an additional good candidate as an identification target in the qRT-PCR technique.
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http://dx.doi.org/10.3390/pathogens10111361DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8618703PMC
October 2021

A rich mosaic of resistance in extended-spectrum β-lactamase-producing Escherichia coli isolated from red foxes (Vulpes vulpes) in Poland as a potential effect of increasing synanthropization.

Sci Total Environ 2021 Nov 20:151834. Epub 2021 Nov 20.

University of Life Sciences, Faculty of Veterinary Medicine, Department of Preclinical Veterinary Sciences, Sub-Department of Veterinary Microbiology, Akademicka 12, 20-033 Lublin, Poland. Electronic address:

In our research, we analyzed the resistance of cephalosporin-resistant E. coli strains to antimicrobial agents. The strains were collected during five years from wild animal species commonly inhabiting Poland. We have identified the type of β-lactamases produced and the multidrug-resistance profile. Most strains (73.8%) had genes encoding ESBL enzymes, mainly CTX-M-1 and TEM. Almost all AmpC-β-lactamase-producing isolates had the bla gene. Almost 70% of the strains tested showed a multi-drug resistance profile. The dominant phenotype was resistance to tetracycline (69.05%), and/or sulfamethoxazole (57.1%). We also found high resistance to quinolones: ciprofloxacin 35.7% and nalidixic acid 52.4%. The phenotypic resistance of the strains was in most cases confirmed by the presence of corresponding genes. Among strains, 26.2% were carriers of plasmid-mediated quinolone resistance genes (PMQR). MLST analysis revealed a large clonal variation of the strains, which was reflected in 28 different sequence types. More than half of the strains (54.7%) were classified into the following sequence complexes: 10, 23, 69, 101, 155, 156, 168, 354, 398, 446, and 648. Only one strain in the studied group was assigned to the ExPEC pathotype and represented sequence type 117. The results of our research have confirmed that isolates obtained from wild animals possess many resistance determinants and sequence types, which are also found in food-producing animals and humans. This reflects the doctrine of "One health", which clearly indicates that human health is inextricably linked with animal health as well as degree of environmental contamination. We conclude that the resistance and virulence profiles of strains isolated from wildlife animals may be a resultant of various sources encountered by animals, creating a rich and varied mosaic of genes, which is very often unpredictable and not reflected in the correlation between the sequence type and the gene profile of resistance or virulence observed in epidemic clones.
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http://dx.doi.org/10.1016/j.scitotenv.2021.151834DOI Listing
November 2021

Virulence and Antimicrobial Resistance Pattern of spp. Colonizing European Pond Turtles and Their Natural Environment. First Study from Poland.

Animals (Basel) 2021 Sep 23;11(10). Epub 2021 Sep 23.

Department of Pediatric Orthopedics and Rehabilitation, Faculty of Medicine, Medical University, Gębali 6, 20-093 Lublin, Poland.

The aim of the study was to isolate and identify species belonging to the genus and evaluate the antimicrobial resistance and virulence patterns of isolates colonizing European pond turtles () from natural environment of Eastern Poland. In total, 74 turtles and 15 samples of water from their natural environment were examined. More than 40 strains were isolated and identified: ( = 1), ( = 13), ( = 2), ( = 3), and ( = 23). The highest incidence of resistance was noted for ampicillin (100%) and sulfamethoxazole (62.0%), followed by erythromycin and colistin (both 40.5%). Moreover, eight strains were intermediately resistant to meropenem (19%). Most isolates were found to possess more than one virulence gene among A, , and . We showed that the population of free-living European pond turtles was highly colonized by spp. Such strains may be an infectious agent not only for the population of turtles but also for other species of animals inhabiting their natural environment. Moreover, the undesirable properties of water quality caused by the presence of drug-resistant aeromonads could have a negative impact on human health.
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http://dx.doi.org/10.3390/ani11102772DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8532793PMC
September 2021

European Hedgehogs (Erinaceus europaeus L.) as a Reservoir of Dermatophytes in Poland.

Microb Ecol 2021 Sep 18. Epub 2021 Sep 18.

Department of Veterinary Microbiology, Faculty of Veterinary Medicine, University of Life Sciences, Akademicka 12, 20-033, Lublin, Poland.

The European hedgehog (Erinaceus europaeus Linnaeus) frequently colonises areas located close to human life in cities, as these are more suitable nest sites offering an abundance of food and allowing avoidance of predators. However, urbanisation has a significant impact on the epidemiology of infectious diseases, including dermatophytoses, the primary source of which are wild animals. In this study, we determined the spectrum of dermatophytes isolated from the European hedgehog and assessed their susceptibility profile to antifungal drugs. Symptomatic and asymptomatic dermatophyte infections were observed in 7.7% and 8% of the 182 examined free-living hedgehogs, respectively. In the pool of the isolated dermatophyte strains, Trichophyton erinacei was dominant (29.9%), followed by Trichophyton mentagrophytes (17.9%), Trichophyton benhamiae (13.4%), Nannizzia gypsea (11.9%), Microsporum canis (10.4%), Nannizzia nana (7.5%), Paraphyton cookei (6.0%), and Nannizzia fulva (3.0%). Susceptibility tests revealed the highest activity of luliconazole and the lowest of activity fluconazole among the azole drugs applied. Although terbinafine generally exhibited high efficacy, two Trichophyton mentagrophytes isolates showed resistance to this drug (MIC = 2 µg/ml) resulting from missense mutations in the SQLE gene corresponding to the amino acid substitution Leu393Phe. Summarising, our study has also revealed that such wildlife animals as hedgehogs can be a reservoir of pathogenic human dermatophytes, including harmful strains resistant to commonly used antifungal drugs.
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http://dx.doi.org/10.1007/s00248-021-01866-wDOI Listing
September 2021

Cold atmospheric pressure plasma (CAPP) as a new alternative treatment method for onychomycosis caused by Trichophyton verrucosum: in vitro studies.

Infection 2021 Dec 9;49(6):1233-1240. Epub 2021 Sep 9.

Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Institute of Preclinical Veterinary Sciences, University of Life Sciences, Akademicka 12, 20-033, Lublin, Poland.

Purpose: Anthropophilic dermatophytes as etiological factors of onychomycoses are more common than zoophilic fungi. In the case of the latter, reverse zoonoses are possible, which poses a threat to the persistence of dermatophytes in the environment. Nevertheless, without treatment, both types of tinea unguium may lead to complete nail plate destruction and secondary mixed infections with fungi and bacteria. One of the zoophilic dermatophytes that cause onychomycosis is Trichophyton verrucosum, whose prevalence has been increasing in recent years. Such infections are usually treated with allylamines and/or azoles, but such a conventional treatment of infections caused by T. verrucosum often fails or is discontinued by patients.

Methods: Herein, we reveal the results of our in vitro studies related to direct application of cold atmospheric pressure plasma (CAPP) on Trichophyton verrucosum growth, germination and adherence to nail as a new alternative treatment method of such types of dermatomycoses.

Results: Our in vitro studies showed that, while exposure to CAPP for 10 min delays germination of conidia and clearly impairs the fitness of the fungal structures, 15 min is enough to kill all fungal elements exposed to plasma. Moreover, the SEM images revealed that T. verrucosum cultures exposed to CAPP for 10 and 15 min were not able to invade the nail fragments.

Conclusion: The results revealed that single exposure to CAPP was able to inhibit T. verrucosum growth and infection capacity. Hence, cold atmospheric pressure plasma should be considered as a promising alternative treatment of onychomycoses.
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http://dx.doi.org/10.1007/s15010-021-01691-wDOI Listing
December 2021

Modulation of ERG gene expression in fluconazole-resistant human and animal isolates of Trichophyton verrucosum.

Braz J Microbiol 2021 Dec 5;52(4):2439-2446. Epub 2021 Aug 5.

Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Institute of Preclinical Veterinary Sciences, University of Life Sciences, Akademicka 12, 20-033, Lublin, Poland.

Dermatophytes are a group of eukaryotic microorganisms characterized by high capacity to colonize keratinized structures such as the skin, hair, and nails. Over the past years, the incidence of infections caused by zoophilic species, e.g., Trichophyton verrucosum, has been increasing in some parts of the world, especially in Europe. Moreover, the emergence of recalcitrant dermatophytoses and in vitro resistant dermatophytes has become a cause of concern worldwide. Here, we analyzed the mechanisms underlying resistance to fluconazole among clinical isolates of T. verrucosum. Quantitative RT-PCR was carried out to determine the relative expression levels of mRNA transcripts of ERG3, ERG6, and ERG11 genes in the fungal samples using the housekeeping gene GAPDH as a reference. Our results showed that the upregulation of the ERG gene expression is a possible mechanism of resistance to fluconazole in this species. Furthermore, ERG11 is the most statistically significantly overexpressed gene in the pool of fluconazole-resistant T. verrucosum isolates. Additionally, we have demonstrated that exposure to fluconazole increases the levels of expression of ERG genes in fluconazole-resistant isolates of T. verrucosum. In conclusion, this study has shown one of the possible mechanisms of resistance to fluconazole among zoophilic dermatophytes, which involves the maintenance of high levels of expression of ERG genes after drug exposure.
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http://dx.doi.org/10.1007/s42770-021-00585-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8578519PMC
December 2021

Real-Time PCR as an Alternative Technique for Detection of Dermatophytes in Cattle Herds.

Animals (Basel) 2021 Jun 2;11(6). Epub 2021 Jun 2.

Department of Veterinary Microbiology, Faculty of Veterinary Medicine, University of Life Sciences in Lublin, Akademicka 12, 20-033 Lublin, Poland.

Dermatophytes are filamentous fungi with the ability to digest and grow on keratinized substrates. The ongoing improvements in fungal detection techniques give new scope for clinical implementations in laboratories and veterinary clinics, including the monitoring of the disease and carrier status. The technologically advanced methods for dermatophyte detection include molecular methods based on PCR. In this context, the aim of this study was to carry out tests on the occurrence of dermatophytes in cattle herds using qPCR methods and a comparative analysis with conventional methods. Each sample collected from ringworm cases and from asymptomatic cattle was divided into three parts and subjected to the real-time PCR technique, direct light microscopy analysis, and culture-based methods. The use of the real-time PCR technique with pan-dermatophyte primers detected the presence of dermatophytes in the sample with a 10.84% (45% vs. 34.17%) higher efficiency than direct analysis with light microscopy. Moreover, a dermatophyte culture was obtained from all samples with a positive qPCR result. In conclusion, it seems that this method can be used with success to detect dermatophytes and monitor cowsheds in ringworm cases and carriers in cattle.
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http://dx.doi.org/10.3390/ani11061662DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8229919PMC
June 2021

In vitro evaluation of photodynamic activity of methylene blue against Trichophyton verrucosum azole-susceptible and -resistant strains.

J Biophotonics 2021 10 7;14(10):e202100150. Epub 2021 Jul 7.

Faculty of Veterinary Medicine, Department of Veterinary Microbiology, University of Life Sciences, Lublin, Poland.

The intense search for the "Holy Grail" of antifungal therapy can be observed today. The searches are not limited only to discovery of potential antifungal drugs, but also new therapeutic strategies involving the use of chemosensitizers to achieve synergistic effect or physicochemical factors inducing stress conditions in fungal cells. In this study was examined in vitro effectiveness of photodynamic antifungal strategy with methylene blue using a light beam with a wavelength equal to 635 nm toward the Trichophyton verrucosum susceptible and itraconazole- and/or fluconazole-resistant strains. Methylene blue used at concentration equal to 5 μg/mL and in the presence of 40 J/cm of light energy showed fungicidal effect toward the susceptible strains. However, for azole-resistant isolates, only the energy dose equal to 60 J/cm at 5 μg/mL of methylene blue allowed to kill the pathogen. This study confirms that methylene blue induced by red light has a definite inhibitory effect on zoophilic dermatophytes.
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http://dx.doi.org/10.1002/jbio.202100150DOI Listing
October 2021

Effect-directed analysis as a method for quality and authenticity estimation of Rhodiola rosea L. preparations.

J Chromatogr A 2021 Jul 2;1649:462217. Epub 2021 May 2.

Department of Chromatography, Institute of Chemical Sciences, Faculty of Chemistry, Maria Curie-Skłodowska University, Maria Curie-Skłodowska sq.3, 20-031 Lublin, Poland. Electronic address:

Adulterations of food and pharmaceutical preparations are the important global problem. On the one hand, fraud practices are becoming more and more sophisticated while on the other, monitoring and uncovering falsifications are insufficient. One of the most common consumer concern is the quality and authenticity of the purchased products, related to the confidence that they have composition and properties in accordance with the manufacturer's declaration on the label. This refers also to pharmaceuticals potentially delivering great health benefits such as Rhodiola rosea L. supplements. The aim of this study was defining authenticity and possible adulterations of two R. rosea preparations basing on their TLC-bioprofiles and the presence of biomarker compounds characteristic for this plant. The effect-directed analysis (EDA), i.e. TLC hyphenated with micro-chemical and biological assays performed directly on TLC plates followed by HPLC-ESI-MS was used for the bioprofiling of antioxidants, antibacterials, and inhibitors of lipase, acetylcholine, α-glucosidase and tyrosinase as well as for the identification of the biomarkers. The results pointed to the possible adulteration of one of the tested products related to the absence of two rosavins, the most important quality markers of R. rosea.
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http://dx.doi.org/10.1016/j.chroma.2021.462217DOI Listing
July 2021

Dataset of the next-generation sequencing of variable 16S rRNA from bacteria and ITS2 regions from fungi and plants derived from honeybees kept under anthropogenic landscapes.

Data Brief 2021 Jun 2;36:107019. Epub 2021 Apr 2.

School of Biological and Chemical Sciences, Queen Mary University of London, London E1 4NS, United Kingdom.

Forager honeybees were collected from four localities located in Europe, i.e.: London, UK; Athens, Greece; Marchamalo, Spain and Lublin, Poland. Furthermore, from Asia we have collected as well as foragers form Chiang Mai in Thailand We used next generation sequencing (NGS) to analyse the 16S rRNA bacterial gene amplicons based on the V3-V4 region and the ITS2 region from fungi and plants derived from honeybee samples. Amplicon libraries, were prepared using the 16S Metagenomic Sequencing Library Preparation, Preparing 16S Ribosomal RNA Gene Amplicons for the Illumina MiSeq System (Illumina®) protocol. NGS raw data are available at https://www.ncbi.nlm.nih.gov/bioproject/PRJNA686953. Furthermore, isolated DNA was used as the template for screening pathogens: , tracheal mite (), any organism in the parasitic order Trypanosomatida, including spp. (i.e., ), neogregarines including and spp. (i.e., ). The presented data can be used to compare the metagenomic samples from different honeybee population all over the world. A higher load of fungi, and bacteria groups such as: Firmicutes (); proteobacteria Neisseriaceae and other unidentified bacteria was observed for and neogregarines infected honeybees. Healthy honeybees had a higher load of plant pollens, and bacteria groups such as: and Enterobacteriaceae More details can be found in research article [1] Ptaszyńska et al. 2021.
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http://dx.doi.org/10.1016/j.dib.2021.107019DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079459PMC
June 2021

Therapeutic efficacy of topically used luliconazole vs. terbinafine 1% creams.

Mycoses 2021 Aug 30;64(8):967-975. Epub 2021 Apr 30.

Hollings Cancer Center, Medical University of South Carolina (MUSC), Charleston, SC, USA.

Objectives: Dermatomycoses of zoophilic origin, especially those caused by Trichophyton mentagrophytes, often pose considerable therapeutic problems. This is reflected in the growing number of strains of this species with resistance to terbinafine caused by a mutation in the squalene epoxidase (SQLE) gene. Therefore, it is reasonable to look for alternative therapies to the commonly used terbinafine. The aim of the present study was to assess the in vivo effectiveness of topical therapy with luliconazole or terbinafine 1% cream.

Methods: Therapeutic efficacy was assessed using direct examination in KOH with DMSO, qPCR analysis with pan-dermatophyte primers and culturing. Moreover, in vitro susceptibility tests for luliconazole and terbinafine were performed.

Results: The results demonstrated significantly higher antifungal activity of luliconazole than terbinafine against dermatomycoses caused by T. mentagrophytes. The geometric mean of the MIC value for luliconazole against all T. mentagrophytes strains was 0.002 μg/ml, while this value for terbinafine was 0.004 μg/ml. In all studied cases, 28-day local therapy with luliconazole contributed to complete eradication of the aetiological agent of infection.

Conclusions: Given the increasingly frequent reports of difficult-to-treat dermatophytoses caused by zoophilic terbinafine-resistant strains, the 1% luliconazole cream can be alternative solution in topical therapy.
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http://dx.doi.org/10.1111/myc.13289DOI Listing
August 2021

Amplicon Sequencing of Variable 16S rRNA from Bacteria and ITS2 Regions from Fungi and Plants, Reveals Honeybee Susceptibility to Diseases Results from Their Forage Availability under Anthropogenic Landscapes.

Pathogens 2021 Mar 22;10(3). Epub 2021 Mar 22.

Laboratory of Gene Expression, ECOTECH-Complex, Maria Curie-Sklodowska University, ul. Gleboka 39, 20-612 Lublin, Poland.

European and Asian honeybees are essential crop pollinators. Microbiome studies can provide complex information on health and fitness of these insects in relation to environmental changes, and plant availability. Amplicon sequencing of variable regions of the 16S rRNA from bacteria and the internally transcribed spacer (ITS) regions from fungi and plants allow identification of the metabiome. These methods provide a tool for monitoring otherwise uncultured microbes isolated from the gut of the honeybees. They also help monitor the composition of the gut fungi and, intriguingly, pollen collected by the insect. Here, we present data from amplicon sequencing of the 16S rRNA from bacteria and ITS2 regions from fungi and plants derived from honeybees collected at various time points from anthropogenic landscapes such as urban areas in Poland, UK, Spain, Greece, and Thailand. We have analysed microbial content of honeybee intestine as well as fungi and pollens. Furthermore, isolated DNA was used as the template for screening pathogens: , , , tracheal mite (), any organism in the parasitic order Trypanosomatida, including Crithidia spp. (i.e., ), neogregarines including and spp. (i.e., ). We conclude that differences between samples were mainly influenced by the bacteria, plant pollen and fungi, respectively. Moreover, honeybees feeding on a sugar based diet were more prone to fungal pathogens () and neogregarines. In most samples sp. and neogregarines parasitized the host bee at the same time. A higher load of fungi, and bacteria groups such as Firmicutes (); -proteobacteria, Neisseriaceae, and other unidentified bacteria was observed for and neogregarine infected honeybees. Healthy honeybees had a higher load of plant pollen, and bacteria groups such as: , , , and Enterobacteriaceae. Finally, the period when honeybees switch to the winter generation (longer-lived forager honeybees) is the most sensitive to diet perturbations, and hence pathogen attack, for the whole beekeeping season. It is possible that evolutionary adaptation of bees fails to benefit them in the modern anthropomorphised environment.
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http://dx.doi.org/10.3390/pathogens10030381DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8004708PMC
March 2021

Comparative study of multidrug-resistant obtained from different hosts.

J Med Microbiol 2021 Mar 22;70(3). Epub 2021 Mar 22.

Department of Pharmaceutical Microbiology, Medical University of Lublin, Chodźki 1, 20-093 Lublin, Poland.

The possible transfer of antimicrobial resistance genes between isolates from humans and different animal species, including those not covered by monitoring programs (e.g. pet and wildlife), poses a serious threat to public health. Little is known about occurrence and mechanisms of phenomenon of multidrug resistance of isolated from various host species in Poland. The aim of the study was to characterize multidrug-resistant isolated from humans and animals (livestock, pets and wildlife) in terms of the occurrence of genetic markers determining resistance. Bacterial isolates were tested for phenotypic resistance and the presence of genes encoding resistance to macrolides, tetracycline, aminoglycosides, aminocyclitols and phenicols as well as efflux pump (A), resolvase (X) and integrase () genes. The quinolone resistance-determining regions of A and C were sequenced. Human isolates of were characterized by high-level resistance to: ciprofloxacin, enrofloxacin, erythromycin (100 %), as well, as aminoglycosides resistance (kanamycin - 100%, streptomycin - 78 %, gentamicin - 78%). Regardless of the animal species, high level of resistance of to tetracycline (from 88-100 %), erythromycin (from 82-94 %) and kanamycin (from 36-100 %) was observed. All isolates from wildlife were resistant to fluoroquinolones. However, full susceptibility to vancomycin was observed in all isolates tested. Phenotypic antimicrobial resistance of was identified in the presence of the following resistance genes: (B) (70%), (A) (50 %), (L) (35 %), (K) (34 %), (M) (76 %), (25%), (31%), (68 %), (X) (23 %), and integrase gene () (34 %). A correlation between an amino acid substitution at positions 83 and 87 of A and position 80 of C and the high-level fluoroquinolone resistance in has been observed as well. The level and range of antimicrobial resistance and the panel of resistance determinants is comparable between isolates, despite host species.
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http://dx.doi.org/10.1099/jmm.0.001340DOI Listing
March 2021

Genetic Predisposition and its Heredity in the Context of Increased Prevalence of Dermatophytoses.

Mycopathologia 2021 May 1;186(2):163-176. Epub 2021 Feb 1.

Faculty of Veterinary Medicine, Institute of Preclinical Veterinary Sciences, Department of Veterinary Microbiology, University of Life Sciences, Akademicka 12, 20-033, Lublin, Poland.

Dermatophytosis is a widespread disease with high prevalence and a substantial economic burden associated with costs of treatment. The pattern of this infectious disease covers a wide spectrum from exposed individuals without symptoms to those with acutely inflammatory or non-inflammatory, chronic to invasive, and life-threatening symptoms. Moreover, the prevalence of cutaneous fungal infections is not as high as might be expected. This curious disparity in the dermatophyte infection patterns may suggest that there are individual factors that predispose to infection, with genetics as an increasingly well-known determinant. In this review, we describe recent findings about the genetic predisposition to dermatophyte infections, with focus on inheritance in families with a high frequency of dermatophyte infections and specific host-pathogen interactions. The results of studies indicating a hereditary predisposition to dermatophytoses have been challenged by many skeptics suggesting that the varied degree of pathogenicity and the ecological diversity of this group of fungi are more important in increasing sensitivity. Nonetheless, a retrospective analysis of the hereditary propensity to dermatophytoses revealed at least several proven genetic relationships such as races, CARD9 deficiency, HLA-DR4 and HLA-DR8 type and responsible genes encoding interleukin-22, β-defensin 2 and 4 as well as genetic defects in dectin-1, which increased the prevalence of the disease in families and were involved in the inheritance of the proneness in their members. In future, the Human Genome Diversity Project can contribute to elucidation of the genetic predisposition to dermatophytoses and provide more information.
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http://dx.doi.org/10.1007/s11046-021-00529-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106586PMC
May 2021

Unusual Penile Prolapse with an Infectious Background Caused by the Burkholderia cepacia Complex in a Stallion.

J Equine Vet Sci 2021 02 16;97:103353. Epub 2020 Dec 16.

University of Life Sciences, Faculty of Veterinary Medicine, Institute of Preclinical Veterinary Sciences, Department of Veterinary Microbiology, Lublin, Poland.

Penile prolapse is a disease manifested by an inability to retract the penis into the preputial sheath. It is reported in a variety of animal species, especially in young and intact males. However, penile prolapse in horses is commonly caused by trauma, sexual activity, pseudohermaphroditism, or neurological deficits, and less often by an infectious background. The present case report aimed to determine the etiological factor of penis infection associated with penile prolapse in a stallion in Poland. Our report indicates that the infectious background of penile prolapse was related to the Burkholderia cepacia complex. Based on antibiotic susceptibility results, the stallion was administered effective streptomycin and enrofloxacin treatment and recovered without complications. The following options are likely to be the infection source: contamination of hay or animal hygiene products. Finally, given its ability to grow in antiseptic solutions, difficulties in culturing, and innate multidrug resistance, this microorganism is currently a challenge to both detection and treatment in veterinary medicine cases.
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http://dx.doi.org/10.1016/j.jevs.2020.103353DOI Listing
February 2021

Complementary effect of mechanism of multidrug resistance in Trichophyton mentagrophytes isolated from human dermatophytoses of animal origin.

Mycoses 2021 May 28;64(5):537-549. Epub 2021 Jan 28.

Faculty of Veterinary Medicine, Department of Veterinary Microbiology, Institute of Preclinical Veterinary Sciences, University of Life Sciences, Lublin, Poland.

Background: Dermatophytoses have gained interest worldwide due to the increased resistance to terbinafine and azoles and difficulty in management of these refractory diseases.

Objectives: In this study, we identified and analysed Trichophyton mentagrophytes clinical isolates obtained from humans with infections of animal origin.

Methods: We used quantitative real-time PCR (qRT-PCR) to examine the transcriptional modulation of three MDR genes (PDR1, MDR2 and MDR4) and analysed squalene epoxidase (SQLE) gene sequences from multidrug-resistant Trichophyton mentagrophytes isolates.

Results: The expression profile revealed a 2- to 12-fold increase in mRNA accumulation in the presence of any of the antifungals, compared to cells incubated without drugs. A statistically significant relationship between the isolates exposed to itraconazole and increased expression of the tested genes was revealed. Substantially lower transcription levels were noted for cells exposed to luliconazole, that is, a third-generation azole. Additionally, in the case of 50% of terbinafine-resistant strains, Leu397Phe substitution in the SQLE gene was detected. Furthermore, the reduced susceptibility to itraconazole and voriconazole was overcome by milbemycin oxime.

Conclusions: In conclusion, our study shed more light on the role of the ABC transporter family in T. mentagrophytes, which, if overexpressed, can confer resistance to single azole drugs and even cross-resistance. Finally, milbemycin oxime could be an interesting compound supporting treatment with azole drugs in the case of refractory dermatomycoses.
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http://dx.doi.org/10.1111/myc.13242DOI Listing
May 2021

Last Call for Replacement of Antimicrobials in Animal Production: Modern Challenges, Opportunities, and Potential Solutions.

Antibiotics (Basel) 2020 Dec 9;9(12). Epub 2020 Dec 9.

Department of Pediatric Orthopedics and Rehabilitation, Faculty of Medicine, Medical University, Gębali 6, 20-093 Lublin, Poland.

The constant market demand for easily available and cheap food of animal origin necessitates an increasing use of antibiotics in animal production. The alarming data provided by organizations monitoring drug resistance in indicator and pathogenic bacteria isolated from humans and animals indicate a possible risk of a return to the preantibiotic era. For this reason, it seems that both preventive and therapeutic measures, taken as an alternative to antimicrobials, seem not only advisable but also necessary. Nevertheless, the results of various studies and market analyses, as well as difficulties in the implementation of alternative substances into veterinary medicine, do not guarantee that the selected alternatives will completely replace antimicrobials in veterinary medicine and animal production on a global scale. This publication is a brief overview of the drug resistance phenomenon and its determinants, the steps taken to solve the problem, including the introduction of alternatives to antimicrobials, and the evaluation of some factors influencing the potential implementation of alternatives in animal production. The review also presents two groups of alternatives, which, given their mechanism of action and spectrum, are most comparable to the effectiveness of antibiotics, as emphasized by the authors.
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http://dx.doi.org/10.3390/antibiotics9120883DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762978PMC
December 2020

Analysis of the occurrence and molecular characteristics of drug-resistant strains of Enterococcus faecalis isolated from the gastrointestinal tract of insectivorous bat species in Poland: A possible essential impact on the spread of drug resistance?

Environ Pollut 2021 Jan 23;269:116099. Epub 2020 Nov 23.

Wrocław University of Environmental and Life Sciences, Faculty of Veterinary Medicine, Department of Pathology, Division of Microbiology, C. K. Norwida 31, 50-375, Wrocław, Poland.

Bats are poorly understood as a reservoir of multidrug-resistant strains; therefore, the aim of this study was to determine molecular characterization of multidrug-resistant Enterococcus strains isolated from bat species from Poland. A multi-stage analysis based on targeted isolation of drug-resistant strains (selective media with tetracycline, chloramphenicol, gentamicin, streptomycin, and vancomycin), determination of the phenotypic profile of drug-susceptibility using the disc diffusion method, and amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting) was used for the isolation and differentiation of strains. The applied strategy finally allowed identification of E. faecalis resistant to at least one antimicrobial in 47.2% of the single-animal group and in 46.9% of the pooled samples of bat's guano. Out of the 36 distinct isolates, 69% met the criteria of multi-drug resistance, with a dominant combination of resistance to tetracycline, erythromycin, and rifampicin. Simultaneously, 41.6% of the strains were high-level aminoglycoside resistant (HLAR). In most strains, phenotypic resistance was reflected in the presence of at least one gene encoding resistance to a given drug. Moreover, our research results show that some genes were detected simultaneously in the same strain statistically significantly more frequently. This may confirm that the spread of some genes (tetM and ermB or aph (3')-IIIa as well as gelE and aac (6')-Ie-aph (2″)-Ia or ant (6)-Ia) is associated with their common occurrence on the same mobile genetic element. To our knowledge, this is the first analysis of multidrug-resistance among E. faecalis isolated from bats. Our research demonstrates that the One Health concept is not associated exclusively with food-producing animals and humans, but other species of wildlife animals should be covered by monitoring programs as well. We confirmed for the first time that bats are an important reservoir of multi-resistant E. faecalis strains and could have a great impact on environmental resistance.
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http://dx.doi.org/10.1016/j.envpol.2020.116099DOI Listing
January 2021

A case of anti- pityriasis versicolor therapy that preserves healthy mycobiome.

BMC Dermatol 2020 09 29;20(1). Epub 2020 Sep 29.

Department of Genetics and Biochemistry, Clemson University, Clemson, SC, USA.

Background: The impact of Malassezia yeasts on skin mycobiome and health has received considerable attention recently. Pityriasis versicolor (PV), a common dermatosis caused by Malassezia genus worldwide, is a manifestation of dysbiosis. PV can be associated with hyper- and/or hypopigmented skin lesions. This disease entity is characterized by high percentage of relapses, which demands a proper antifungal therapy that is based on unambiguous species identification and drug susceptibility testing.

Case Presentation: Comprehensive analysis of PV case in man presenting simultaneously hyper- and hypopigmented skin lesions was performed. Conventional and molecular diagnostic procedures revealed Malassezia furfur and Malassezia sympodialis, respectively as etiological agents of skin lesions observed. Susceptibility tests showed significantly lowered sensitivity of M. furfur cells to fluconazole. Based on susceptibility profiles local antifungal therapy with drugs characterized by entirely different mechanism of action was included.

Conclusions: Our study indicates that cases of PV represented by two types of skin lesions in one patient may be associated with distinct Malassezia species. Moreover, as observed in this case, each of the isolated etiological agents of PV may differ significantly in susceptibility to antifungals. This can significantly complicate the treatment of dermatosis, which by definition is associated with a significant percentage of relapses. In the presented case localized topical treatment was sufficient and successful while allowing maintaining the physiological mycobiome.
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http://dx.doi.org/10.1186/s12895-020-00106-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7526128PMC
September 2020

Wildlife omnivores and herbivores as a significant vehicle of multidrug-resistant and pathogenic Escherichia coli strains in environment.

Environ Microbiol Rep 2020 12 28;12(6):712-717. Epub 2020 Sep 28.

Sub-Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Institute of Preclinical Veterinary Sciences, University of Life Sciences, Akademicka 12, Lublin, 20-033, Poland.

The phenomenon of resistance of Escherichia coli strains in free-living animals has been constantly expanding in recent years. However, the data are still fragmented and, due to the growing threat to public health, there is a constant need to search for and analyse new reservoirs and indicate their role and importance in the circulation of resistance genes in the environment. Therefore, the target group in this study were free-living non-predatory animals as reservoirs of drug-resistant and potentially virulent E. coli strains. We obtained 70 different isolates, including 71.4% of multidrug-resistant strains. In strains isolated from all species of animals, we determined high resistance to ampicillin (95.7%), tetracycline (64.3%), streptomycin (51.4%) and chloramphenicol (38.6%). Every third of the E. coli-positive individual was a carrier of more than one resistant clone. Moreover, 11.4% of isolates among the resistant strains had the ExPEC, ETEC, or EHEC pathotype. Our study confirmed that not only free-living predatory animals are reservoirs of resistance but also many synanthropic species of herbivores and omnivores contribute substantially to the spread of resistant and virulent E. coli strains.
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http://dx.doi.org/10.1111/1758-2229.12886DOI Listing
December 2020

Wildlife Carnivorous Mammals As a Specific Mirror of Environmental Contamination with Multidrug-Resistant Strains in Poland.

Microb Drug Resist 2020 Sep 28;26(9):1120-1131. Epub 2020 Apr 28.

Sub-Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Institute of Biological Bases of Animal Diseases, University of Life Sciences, Lublin, Poland.

In recent decades, the number of studies on the occurrence of resistant strains in wildlife animals has increased significantly, but data are still fragmentary. The aim of this study was to evaluate drug resistance of strains isolated from wild carnivorous mammals, common in Poland. Selective media with antimicrobials (tetracycline, kanamycin, chloramphenicol, and cefotaxime) were used for isolation. Of 53 isolates shown to be distinct by the amplification of DNA fragments surrounding rare restriction site-fingerprinting method, 77.8% were multidrug-resistant (multidrug-resistant). All strains were resistant to ampicillin and many of them also exhibited resistance to tetracycline (76.2%), sulfamethoxazole (57.1%), streptomycin and kanamycin (49.2%), chloramphenicol (30.1%), and nalidixic acid (46%). In most cases, the phenotypic resistance profile was confirmed by detection of relevant genes mostly occurring in strains isolated from livestock animals and humans. Extended-spectrum β-lactamase-producing strains were detected in one mink and three martens. The strains were carriers of , and genes. Our research confirmed a high carrier rate of MDR , even more than one MDR strain in a single individual; therefore, wider monitoring in this group of animals should be considered.
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http://dx.doi.org/10.1089/mdr.2019.0480DOI Listing
September 2020

Dermatophytosis with concurrent Trichophyton verrucosum and T. benhamiae in calves after long-term transport.

Vet Dermatol 2020 Oct 26;31(5):414-e111. Epub 2020 Aug 26.

State Veterinary Laboratory, Słowicza 2, Lublin, 20-336, Poland.

Background: Dermatophytosis is a common problem in cattle. The aetiological factors associated with this disease are filamentous fungi with the ability to digest and grow on keratinized substrates. In cattle, and less frequently in other domestic animals and people, the dermatophyte Trichophyton verrucosum is most commonly isolated from skin lesions. The dermatophyte Trichophyton benhamiae is an important zoonotic pathogen, and the main sources of transmission are guinea pigs and other small rodents.

Objectives: In this report, we show multispecies infection in calves (Bos taurus) after long-term transport and vaccination against trichophytosis.

Animals: Sixty animals were imported of which 32 were observed to be affected with superficial infection nine to 12 days after vaccination for dermatophytosis.

Methods And Materials: Diagnosis was made correlating the clinical signs with a micro- and macroscopic examination of cultured fungi. Molecular differentiation was used to confirm the species affiliation.

Results: Eight of the calves were infected with T. verrucosum alone, and 24 calves with both T. verrucosum and T. benhamiae. We suggest that the cause of this large outbreak was immunosuppression of the animals resulting from the stress of transport and administration of vaccine.

Conclusion: Both T. verrucosum and T. benhamiae can be seen concurrently in cattle.
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http://dx.doi.org/10.1111/vde.12880DOI Listing
October 2020

Intrinsic resistance to terbinafine among human and animal isolates of Trichophyton mentagrophytes related to amino acid substitution in the squalene epoxidase.

Infection 2020 Dec 8;48(6):889-897. Epub 2020 Aug 8.

Department of Mycology and Genetics, Faculty of Biological Sciences, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland.

Background: Dermatomycoses are the most common fungal infections in the world affecting a significant part of the human and animal population. The majority of zoophilic infections in humans are caused by Trichophyton mentagrophytes. Currently, the first-line drug for both oral and topical therapy is terbinafine. However, an increasing number of cases that are difficult to be cured with this drug have been noted in Europe and Asia. Resistance to terbinafine and other allylamines is very rare and usually correlated with point mutations in the squalene epoxidase gene resulting in single amino acid substitutions in the enzyme, which is crucial in the ergosterol synthesis pathway.

Purpose: Here, we report terbinafine-resistant T. mentagrophytes isolates among which one was an etiological factor of tinea capitis in a man and three were obtained from asymptomatic foxes in Poland.

Methods: We used the CLSI protocol to determine antifungal susceptibility profiles of naftifine, amphotericin B, griseofulvin, ketoconazole, miconazole, itraconazole, voriconazole, and ciclopirox. Moreover, the squalene epoxidase gene of the terbinafine-resistant strains was sequenced and analysed.

Results: In the genomes of all four resistant strains exhibiting elevated MICs to terbinafine (16 to 32 µg/ml), single-point mutations leading to Leu393Phe substitution in the squalene epoxidase enzyme were revealed. Among the other tested substances, a MIC50 value of 1 µg/ml was shown only for griseofulvin.

Conclusion: Finally, our study revealed that the terbinafine resistance phenomenon might not be acquired by exposure to the drug but can be intrinsic. This is evidenced by the description of the terbinafine-resistant strains isolated from the asymptomatic animals.
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http://dx.doi.org/10.1007/s15010-020-01498-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7674369PMC
December 2020

Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals.

Eur J Clin Microbiol Infect Dis 2020 Nov 30;39(11):2099-2113. Epub 2020 Jun 30.

Faculty of Agrobioengineering, Department of Management and Marketing, University of Life Sciences, Dobrzanskiego 37, 20-626, Lublin, Poland.

The major problems in determining the causative factors of the high prevalence of dermatophytoses include the lack of a well-standardized antifungal susceptibility testing method, the low consistency of in vitro and clinical minimal inhibitory concentration values, the high genomic diversity of the population, and the unclear mechanism of pathogenicity. These factors are of particular importance when the disease is recalcitrant and relapses. Herein, we identified and characterized Trichophyton mentagrophytes isolates obtained from therapy-resistant cases in humans and animals. We used genomic diversity analysis of 17 human and 27 animal clinical isolates with the MP-PCR technique, determined their phenotypic enzymatic activity and host range, and performed antifungal susceptibility testing to currently available antifungal drugs from various chemical groups. Genomic diversity values of 35.3% and 33.3% were obtained for clinical isolates from humans and animals, respectively, yet without any relationship to the host species or antifungal drug to which resistance in therapy was revealed. The highest activity of keratinase enzymes was recorded for fox, guinea pig, and human hairs. These hosts can be considered as the main species in the host range of these isolates. A phenyl morpholine derivative, i.e. amorolfine, exhibited superior activity against strains obtained from both humans and animals with the lowest MIC. Interestingly, high compliance of terbinafine in vitro resistance with clinical problems in the treatment with this substance was shown as well. The high resistance of dermatophytes to drugs is the main cause of the recalcitrance of the infection, whereas the other features of the fungus are less important.
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http://dx.doi.org/10.1007/s10096-020-03952-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7561545PMC
November 2020

Identification of emerging trends in the prevalence of dermatophytoses in alpacas (Vicugna pacos) farmed in Poland.

Transbound Emerg Dis 2020 Nov 24;67(6):2702-2712. Epub 2020 May 24.

Department of Mycology and Genetics, Faculty of Biological Sciences, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland.

The increasing number of dermatophytoses among animals observed recently in developed countries may be connected with relocation of many exotic species outside their natural living environment. Moreover, an impact on this situation may also be exerted by relapses related to limited compliance with antifungal treatment regimes. Many exotic animals, including camelids, imported to European countries are connected with cases of tuberculosis or zoophilic dermatophytoses in humans. In the present study, we identified and comprehensively analysed dermatophyte infections in alpacas from breeding farms in Poland. As part of this study, we determined the prevalence of dermatophyte infections in alpacas. The conventional and molecular mycological diagnostic procedures applied led to unambiguous identification of the aetiological factors of symptomatic dermatophytoses and asymptomatic animals, that is Trichophyton benhamiae and T. verrucosum. Furthermore, the susceptibility tests allowed choosing the best therapeutic option and revealed superior activity of allylamine drugs against all strains. Finally, in the case of strains isolated from symptomatic dermatophytoses, our study revealed a significantly higher virulence level expressed by high activity of chosen enzymes, especially related to keratinolytic and haemolytic activity. In conclusion, this report indicates that farmed alpacas can be a reservoir or vector for contagious zoophilic dermatophyte infection. For this reason, imported animals should be subjected to meticulous monitoring to detect not only symptomatic infections but also asymptomatic animals.
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http://dx.doi.org/10.1111/tbed.13624DOI Listing
November 2020

Contamination of the urban environment with excrements of companion animals as an underestimated source of Staphylococcus species posing a threat to public health.

Acta Vet Hung 2020 03 8;68(1):12-19. Epub 2020 May 8.

2Department of Genetics and Microbiology, Maria Curie-Skłodowska University, Lublin, Poland.

The aim of the study was to assess the incidence, resistance, virulence, and genotypic characteristics of Staphylococcus spp. residing in the gastrointestinal tract of dogs and cats, as a group of animals causing potential contamination of the urban space. A high percentage of strains resistant to penicillin (58%), oxacillin (9%) and tetracycline (60%) were found. All isolates resistant to penicillin, kanamycin, or chloramphenicol carried genes responsible for individual resistance (blaZ, aph(3')-IIIa, and cat (pC194)/cat (pC223), respectively. The mecA gene was detected in 45% of the oxacillin-resistant Staphylococcus pseudintermedius strains. The amplification of DNA fragments surrounding rare restriction sites analysis demonstrated high heterogeneity of genotypic profiles correlating with phenotypic resistance profiles. Multilocus sequence typing analysis classified the methicillin-resistant S. pseudintermedius strains as ST71, ST890, and the totally new ST1047. The presence of a high level of resistance among Staphylococcus strains may suggest a potential risk of transfer of these bacteria between companion animals and humans.
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http://dx.doi.org/10.1556/004.2020.00018DOI Listing
March 2020

Unusual dermatomycoses caused by Nannizzia nana: the geophilic origin of human infections.

Infection 2020 Jun 30;48(3):429-434. Epub 2020 Mar 30.

Department of Mycology and Genetics, Faculty of Biological Sciences, Institute of Genetics and Microbiology, University of Wroclaw, Przybyszewskiego 63/77, 51-148, Wroclaw, Poland.

Background: Fungal infections of the skin, hair, and nails are the largest and most widespread group of all mycoses. Nannizzia nana is a relatively rare etiological factor of dermatomycosis in humans, as it usually affects animals, e.g. pigs and boars. In addition to the zoophilic nature, there are also reports of the geophilic reservoir of this dermatophyte species.

Objective: In this study, we present symptomatic infections with N. nana aetiology in humans reported recently in Poland. Interestingly, these cases had a non-specific clinical picture and occurred as skin lesions on the neck and foot as well as onychomycosis of the toenails. From the medical history, the patients had no contact with pigs.

Methods: Diagnostics of these infections was performed with a combination of classical phenotypic and molecular genomic methods. The genomic diversity of the isolates was determined using the MP-PCR method. In vitro antifungal susceptibility tests against itraconazole, ketoconazole, terbinafine and naftifine hydrochloride were also performed.

Results: Nannizzia nana has been identified as an etiological factor of dermatomycosis. Moreover, heterogeneity of the genomes was revealed for the obtained strains. In vitro activities of antifungal agents showed that isolates were susceptible to all tested drugs. The patients were treated with oral terbinafine and topical ketoconazole cream, which led to a complete recovery.

Conclusions: In conclusion, the cases studied by us may indicate that the infrequency of N. nana infections may not necessarily be related to the low infectivity of this fungal agent, but they are rather associated with misdiagnosis. Furthermore, N. nana reservoirs should also be sought in soil.
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http://dx.doi.org/10.1007/s15010-020-01416-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256082PMC
June 2020

Effect directed analysis and TLC screening of Schisandra chinensis fruits.

J Chromatogr A 2020 May 3;1618:460942. Epub 2020 Feb 3.

Department of Chromatography, Faculty of Chemistry, University of M. Curie-Skłodowska, M. Curie-Skłodowska Sq. 3, 20-031 Lublin, Poland. Electronic address:

The fruit of Schisandra chinensis (Chinese magnolia vine), the medicinal plant well-known in Traditional Chinese Medicine, gains great popularity in the modern phytopharmacology. This phenomena is related to the wide and powerful healing properties, including supporting immune, nervous and digestive systems activity. S. chinensis is also known for its adaptogenic properties which can support the treatment of neurodegenerative disorders, particularly Alzheimer's disease. The components of S. chinensis have been analyzed mostly using chromatography, including HPLC, GC and TLC. The last technique can be easily hyphenated with biological assays performed directly on a TLC plate that is using effect directed detection. TLC-direct bioautography (TLC-DB) for acetylcholinesterase (AChE) inhibition and TLC-DB against Bacillus subtilis were performed, followed by micro-preparative separation of fractions which were subsequently subjected to LC-MS tentative identification. Additionally, screening analysis was done using both biological detection and derivatization reagents (e.g. PMA, thymol, NP-PEG). Both TLC screening and effect-directed analysis (TLC-DB followed by MS analysis) of S. chinensis fruit revealed components with biological activity, especially antibacterials (e.g. citric acid) and inhibitors of AChE (mainly dibenzocyclooctadiene lignans). AChE inhibition activities were confirmed by TLC-DB for nine standards that is: 6-O-benzoylgomisin, deoxyschisandrin, gomisin A, gomisin G, schisandrin, schisandrin C, schisanhenol, schisantherin A and schisantherin B. These lignans were further identified by LC-ESI-MS in the isolated fraction revealing AChE inhibition. Moreover three other lignans: γ-schisandrin, schisandrin B and gomisin J were tentatively identified by LC-ESI-MS.
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http://dx.doi.org/10.1016/j.chroma.2020.460942DOI Listing
May 2020

Comparison of in vitro activities of 11 antifungal agents against Trichophyton verrucosum isolates associated with a variety hosts and geographical origin.

Mycoses 2020 Mar 23;63(3):294-301. Epub 2019 Dec 23.

Sub-Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Institute of Biological Bases of Animal Diseases, University of Life Sciences, Lublin, Poland.

The high prevalence of dermatophytosis in animals is usually associated with extra expenditure on prevention, diagnosis and long-term treatment. Humans are usually infected from animals, also from asymptomatic carriers, through direct contact or indirectly via fungus-bearing hair, scales and fomites. Despite the medical importance of Trichophyton verrucosum infections, there are limited in vitro data on the fungal susceptibility to antifungal drugs, including new-generation triazoles, imidazoles and allyloamines. The aim of the current study was to evaluate comprehensively the in vitro activity of 11 antifungal drugs against a large collection of T. verrucosum isolates obtained in Poland, Latvia, Lithuania and Slovakia from humans and animals using a microdilution assay. In vitro susceptibility testing of 11 antifungal drugs was performed according to the Clinical and Laboratory Standards Institute (CLSI) document M38. The MICs of clotrimazole, ciclopirox, enilconazole, miconazole, naftifine and terbinafine against all T. verrucosum isolates were below 1 μg/mL, whereas those of fluconazole, griseofulvin, itraconazole, ketoconazole and voriconazole were above 1 μg/mL. Ciclopirox was demonstrated to have superior activity against all strains in comparison with the other drugs, whereas fluconazole exerted the weakest in vitro effect and exhibited the highest MIC values. Our study has shown that drugs of different chemical origin have satisfactory antifungal activity and can be promising candidates for the treatment of T. verrucosum dermatophytosis. Moreover, no significant disparity in drug sensitivity between isolates obtained from different hosts and geographical regions have been demonstrated.
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http://dx.doi.org/10.1111/myc.13042DOI Listing
March 2020

Tinea corporis caused by Trichophyton equinum transmitted from asymptomatic dogs to two siblings.

Braz J Microbiol 2020 Sep 9;51(3):1433-1438. Epub 2019 Dec 9.

Faculty of Biological Sciences, Institute of Genetics and Microbiology, Department of Mycology and Genetics, University of Wroclaw, Przybyszewskiego 63/77, 51-148, Wroclaw, Poland.

Each year, millions of people worldwide are affected by superficial mycoses, which are frequently caused by dermatophytes having affinity to the scalp, nails, hair and the stratum corneum of the skin. The changing reservoirs of zoophilic dermatophytes, the numerous reports of animal carrier status, and the increasing number of pets make the diagnosis difficult, which is usually impossible based only on medical history. Herein we present a case report of tinea corporis caused by Trichophyton equinum in siblings who had no contact with horses. The routine laboratory diagnostic procedures for identification of isolates were based on phenotypic and genotypic characteristics, especially molecular techniques using rDNA internal transcribed spacer sequences. The results showed that both techniques proved to be insufficiently discriminatory to differentiate two closely related species, i.e. Trichophyton equinum and Trichophyton tonsurans. Introduction of a TEF1 sequence analysis to the diagnostic procedures revealed consistent differences between these two species and facilitated unambiguous identification. Interestingly, dogs that could leave the homestead freely were the source of the infection in children. In conclusion, Trichophyton equinum was considered in the past as a strict zoophilic dermatophyte associated with horses and rarely transmitted to humans. This study revealed that this species can have other reservoirs and live in the fur of asymptomatic animals.
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http://dx.doi.org/10.1007/s42770-019-00204-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7455638PMC
September 2020
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