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.

Splenitis in 33 Dogs.

Splenitis is uncommonly reported in dogs. Herein, the authors describe its prevalence, clinical findings and outcomes, histologic patterns, and causes. Splenic samples of dogs diagnosed with splenitis between 2005 and 2013 were collected and stained with hematoxylin and eosin, Gram, green-Gram, Giemsa, periodic acid-Schiff, and Ziehl-Neelsen. Samples were processed for polymerase chain reaction (PCR) to detect bacteria, fungi, and protozoa ( Leishmania infantum, Hepatozoon canis). Thirty-three of 660 splenic samples (5%) had splenitis. Clinical findings and outcomes were available in 19 dogs (58%); 49% had weakness, 33% had fever, and 84% survived. The most frequent inflammatory patterns included purulent splenitis (27%), pyogranulomatous splenitis (24%), and neutrophilic perisplenitis (15%). One dog had a putative diagnosis of primary splenitis; in 8 dogs, microorganisms were identified histologically or by PCR in the spleen without obvious comorbidities. Twenty-four dogs (73%) had concurrent diseases; a permissive role in the development of splenitis was suspected in 21 of these cases. Histologic examination identified the cause of splenitis in 10 dogs. Bacteria were identified by PCR in 23 cases, but the bacteria were confirmed histologically in only 6 of these. Leishmania was detected with PCR in 6 dogs. Leishmania was identified in 1 dog and H. canis in another histologically, but both were PCR negative. Fungi were identified in 8 spleens by PCR and in 1 by histology. This study suggests that splenitis is uncommon in dogs and is frequently associated with systemic diseases. Prognosis is favorable in most cases. Identification of bacteria, fungi, and protozoa in the spleens of affected dogs with PCR should be interpreted cautiously, because the findings are not confirmed histologically in many cases.

Agar Diffusion Procedures for Susceptibility Testing of Malassezia pachydermatis: Evaluation of Mueller-Hinton Agar Plus 2 % Glucose and 0.5 µg/ml Methylene Blue as the Test Medium.

Aim of this study was to verify whether Mueller-Hinton agar supplemented with 2 % glucose and methylene blue (MH-GM), which is used for disk diffusion susceptibility testing of Candida species by the Clinical and Laboratory Standards Institute, is suitable for testing Malassezia pachydermatis. A variant of the disk diffusion procedure utilizing a 9-mm tablet was used to test 31 isolates against clotrimazole and miconazole using MH-GM as test medium. The MH-GM agar optimally supported the growth of all M. pachydermatis isolates, provided that the yeast inoculum was prepared with a lipid source (Tween 40 and 80). Zone edges were frequently definite and clear, facilitating the measurement of zone size and minimizing subjectivity. The inhibition zones correlated with MIC values obtained in a broth dilution assay. The agar diffusion method with MH-GM as the test medium appears as a suitable procedure for testing the susceptibility of M. pachydermatis to CTZ and MCZ in clinical laboratories. This test format may allow processing a large number of isolates in epidemiological studies. This may in turn facilitate clarifying to what extent the problem "drug resistance" accounts for cases of treatment failure in dogs with Malassezia otitis and dermatitis.

In vitro antifungal susceptibility of Malassezia pachydermatis strains isolated from dogs with chronic and acute otitis externa.

Malassezia pachydermatis is a yeast that is frequently involved as a secondary/perpetuating factor in canine otitis externa. Topical therapies with different antifungal agents, mainly azole compounds, are generally successful in controlling the yeast overgrowth, but treatment failure and rapid recurrences are common. This study compared the in vitro antifungal susceptibility of M. pachydermatis isolates obtained from chronic and acute cases of otitis externa. The aim was to assess the possible onset of resistance mechanisms in isolates involved in long-lasting episodes with poor response to treatment. We evaluated the in vitro susceptibility to miconazole (MCZ) and clotrimazole (CTZ) of 42 isolates of M. pachydermatis obtained from dogs with chronic (group A, n = 25) and acute otitis (group B, n = 17), using a modified CLSI M27-A3 microdilution method. All isolates were inhibited by the antifungal agents employed, but Malassezia isolates from group A were significantly associated with higher minimum inhibitory concentration (MIC) values for both agents (Median MIC values: MCZ group A 2 µg/ml, group B 1 µg/ml; CTZ group A 8 µg/ml, group B 4 µg/ml). These findings prove that these isolates had a reduced in vitro susceptibility to the antifungal agents tested. However, it is unlikely that this could have any influence on the outcome of a topical treatment. Indeed, marketed products include concentrations of the tested agents that largely exceed even the highest MICs found in this study (in most cases at least 1,000 × the MIC, or greater). In conclusion, this study suggests that isolates of M. pachydermatis involved in chronic cases of canine external otitis and exposed to repeated antifungal treatments are unlikely to develop mechanisms of resistance of clinical relevance.

Outbreaks of COVID-19 in indoor places of worship: a systematic review.

AIMS: This review aimed to describe what has been published on COVID-19 outbreaks originating from indoor places of worship. METHODS: A systematic review was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist by searching PubMed, Scopus and Embase from 1 January 2020 to 29 March 2021. Citation chasing was also performed. Studies with information about COVID-19 outbreaks originating in indoor places of worship of any religion were included. RESULTS: A total of 9729 records were identified and 36 were selected. The articles reported 119 descriptions of outbreaks linked to churches, mosques, synagogues, and temples, referring to approximately 52-74 unique outbreaks. The outbreaks were mostly located in three major areas: East and Southeast Asia (46%), the USA (27%), Europe (22%). All the outbreaks began in 2020. Mainly, there were no restrictive measures, or such measures were not followed at the time of the outbreak. Choir practices presented the highest attack rate (up to 0.867). CONCLUSIONS: The lack of preventive measures and the role of singing practices were highlighted. Reports were often lacking contact tracing and sometimes did not report the date of outbreak extinction. Moreover, reports came from few geographical areas. Thus, the impact of transmission in places of worship may be largely underestimated.

Oral administration of moxidectin for treatment of murine acariosis due to Radfordia affinis.

A field trial was conducted to assess the safety and efficacy of oral administration of moxidectin in mice naturally infected with the fur mites Radfordia affinis. The natural infection was diagnosed in two colonies within a large academic institution by direct hair examination. Animals received moxidectin (1% Cydectin, FortDodge) at an oral dosage of approximately 2 mg/kg body weight by micropipette; administration was repeated after 15 days. Forty mice served as an untreated control group. Moxidectin treatment resulted in clinical improvement within a few days after initial treatment, and mites were eradicated from all infested animals at day 30. No side effects or signs of ill health were observed in any of the treated animals. To our knowledge, this is the first report of oral moxidectin for treatment of murine acariosis.

Management of Malassezia-related diseases in the dog.

Most cases of Malassezia dermatitis/otitis in the dog are associated with concurrent dermatoses or systemic diseases and recurrences are not uncommon. Recognition and control of the predisposing factors are therefore key factors for successful therapy and prevention of recurrent infections. Currently, Malassezia dermatitis/otitis is managed by the use of antifungal drugs. Systemic therapy is often necessary, in particular when clinical signs are severe and widespread. Ketoconazole and Itraconazole are the most commonly used drugs. Topical therapy is an alternative in case of localized lesions and external ear localizations. Different commercial formulations, available in clinical practice in form of creams, gels, lotions, sprays and ear drops are often used as adiuvants to systemic therapy. Topicals more frequently used are represented by imidazolic antifungals, chlorhexydine and lime sulphur. The presentation deals with more recent advances about the protocols for treatment of Malassezia-related diseases in the dog. New perspectives, as the use of natural compounds, immunotherapy and inhibitors of yeast adherence factors, are also discussed.

A case of disseminated mycosis in a German shepherd dog due to Penicillium purpurogenum.

The genus Penicillium is among the most common contaminant fungi in the environment. Around 15 species are known to cause opportunistic human mycoses, in immunocompromised patients. Until now, Penicillium purpurogenum has been involved in only three human cases of pulmonary diseases but no infections in animals have been reported. Most disseminated mycoses in dogs are caused by members of the genus Aspergillus, with the predisposing factors in these cases being difficult to define. The case reported here involved a 4-year-old female German shepherd dog (GSD) with forelimb instability and back pain. Clinical examination showed hyperthermia, generalized lymphadenomegaly and kyphosis. Radiological findings of the spine revealed areas of discospondilitis involving thoracic and lumbar vertebrae. Microscopic observations of fine needle aspiration biopsies (FNAB) of lymph-nodes showed regular, septate, branching fungal hyphae. Itraconazole therapy was started but the subject died six days later. Disseminated necrotic areas were detected in enlarged lymph-nodes, liver and spleen. Vertebral granulomas within lytic areas in T10-T11 and L2-L3, were observed. Cultures inoculated with samples obtained from lymph-node FNAB and bioptic material from necropsied organs revealed the presence of pure cultures of Penicillium, subsequently identified as P. purpurogenum. Apart from female GSD's suspected predisposition to disseminated mycoses described in literature, no other predisposing factors were ascertained in this case.

Eastern cottontail (sylvilagus floridanus) as carrier of dermatophyte fungi.

Eastern cottontail (Sylvilagus floridanus, fam. Leporidae), introduced into Piedmont (Italy) in the 1960s, was studied as carrier of dermatophyte fungi. Of 216 hair samples collected from animals culled between September 1999 and July 2000 in the Province of Alessandria (Piedmont, Italy) during a pest control project, 57 (26.4%) yielded dermatophyte colonies. As two different species of dermatophytes grew from two samples, a total of 59 fungal isolates (26.5%) were obtained. Six dermatophyte species both geophilic (M. gypseum, M. cookei, Trichophyton ajelloi, T. terrestre) and zoophilic (M. canis, T. mentagrophytes) were identified. No sex-related differences were found but season-related differences were observed. The highest prevalence of dermatophyte-positive samples was recorded in May-September, due to the geophilic fungi whose prevalence decreased during colder and increased during warmer months (p < 0.001). The presence of zoophilic dermatophytes, T. mentagrophytes, commonly associated with rodents, small mammals and lagomorphs and M. canis, usually correlated with domestic environment, did not change the whole year round. As Eastern cottontail has been showed to be a carrier of dermatophytes transmissible to man (M. canis, T. mentagrophytes and M. gypseum), it may represent a source of infection for gamekeepers, hunters and veterinarians.

Seasonal 4-year investigation into the role of the alpine marmot (Marmota marmota) as a carrier of zoophilic dermatophytes.

Two hundred and six samples of alpine marmot (Mamota marmota) hair (148 from adults and 58 from young subjects), 102 soil samples from the entrances to the burrows of the above individuals and 20 control specimens (obtained from adjoining areas away from the burrow systems where the rodents are not usually present) were examined from May 1994 to September 1997. Seventy-five isolates belonging to six species of dermatophytes were found in 69 of the 206 hair samples examined (33.5%). Two species were zoophilic, Microsporum canis (7.8%) and Trichophyton mentagrophytes (11.2.%), and four geophilic, Microsporum cookei (2%), M. gypseum (5.8%), Trichophyton ajelloi (3.9%) and T. terrestre (5.8%). The prevalence of each species in the hair samples did not change significantly according to year, season (chi-squared test [limit significance: P <0.05] gives no significant values [P>0.05] both in year and in season comparison) or age/sex (adult versus juvenile: P=0.1; male versus female: P=0.8) of the marmot. Twenty-three of the 102 soil samples (22.5%) were positive for dermatophytes found in the hair of marmots from the same burrow systems. Five of the 20 control soil samples (25%) were positive for dermatophytes. One isolate of M. gypseum, three of T. terrestre and one of T. mentagrophytes were obtained. Compared with other free-ranging rodent hosts studied in Europe, this mycoflora is characterized by the presence and relatively high prevalence of M. canis, frequently reported in symptomatic and asymptomatic cats, dogs and fur animals. M. canis has not been isolated in other rodents in the wild. However, it has recently been reported in asymptomatic foxes (Vulpes vulpes) from northern Italy. The close link between V vulpes and M. marmota, with the former representing the most important mammal predator of the latter in the Alps (only a fraction of the predator's attacks result in the death of the rodent) may have favoured the adaptation of M. canis to this rodent host. The stable character of the M. canis/M. marmota relationship (no seasonally or annually related difference in the prevalence of this dermatophyte has been found) suggests the inclusion of the alpine marmot in the reservoir of this zoophilic pathogenic agent. In this situation, hibernation in labyrinthine burrow systems, where temperature and moisture ranges are quite uniform the whole year round, may favour the viability of M. canis arthroconidia, whose survival in mountain habitat might otherwise be compromised. This seems to be confirmed by the fact that the fungus has never been found in the control samples collected at a distance of 300 m from the outer edge of the sampled burrow systems.