Amphibian chytridiomycosis : a review with focus on fungus-host interactions

Amphibian declines and extinctions are emblematic for the current sixth mass extinction event. Infectious drivers of these declines include the recently emerged fungal pathogens Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans (Chytridiomycota). The skin disease caused by these fungi is named chytridiomycosis and affects the vital function of amphibian skin. Not all amphibians respond equally to infection and host responses might range from resistant, over tolerant to susceptible. The clinical outcome of infection is highly dependent on the amphibian host, the fungal virulence and environmental determinants. B. dendrobatidis infects the skin of a large range of anurans, urodeles and caecilians, whereas to date the host range of B. salamandrivorans seems limited to urodeles. So far, the epidemic of B. dendrobatidis is mainly limited to Australian, neotropical, South European and West American amphibians, while for B. salamandrivorans it is limited to European salamanders. Other striking differences between both fungi include gross pathology and thermal preferences. With this review we aim to provide the reader with a state-of-the art of host-pathogen interactions for both fungi, in which new data pertaining to the interaction of B. dendrobatidis and B. salamandrivorans with the host's skin are integrated. Furthermore, we pinpoint areas in which more detailed studies are necessary or which have not received the attention they merit

Efficacy of chemical disinfectants for the containment of the salamander chytrid fungus Batrachochytrium salamandrivorans

The recently emerged chytrid fungus Batrachochytrium salamandrivorans (Bsal) causes European salamander declines. Proper hygiene protocols including disinfection procedures are crucial to prevent disease transmission. Here, the efficacy of chemical disinfectants in killing Bsal was evaluated. At all tested conditions, Biocidal (R), Chloramine-T (R), Dettol medical (R), Disolol (R), ethanol, F10 (R), Hibiscrub (R), potassium permanganate, Safe4 (R), sodium hypochlorite, and Virkon S (R), were effective at killing Bsal. Concentrations of 5% sodium chloride or lower, 0.01% peracetic acid and 0.001-1% copper sulphate were inactive against Bsal. None of the conditions tested for hydrogen peroxide affected Bsal viability, while it did kill Batrachochytrium dendrobatidis (Bd). For Bsal, enzymatic breakdown of hydrogen peroxide by catalases and specific morphological features (clustering of sporangia, development of new sporangia within the original sporangium), were identified as fungal factors altering susceptibility to several of the disinfectants tested. Based on the in vitro results we recommend 1% Virkon S (R), 4% sodium hypochlorite and 70% ethanol for disinfecting equipment in the field, lab or captive setting, with a minimal contact time of 5 minutes for 1% Virkon S (R) and 1 minute for the latter disinfectants. These conditions not only efficiently target Bsal, but also Bd and Ranavirus

Bloodstream yeast infections: a 15-month survey

A 15-month survey of 412 bloodstream yeast isolates from 54 Belgian hospitals was undertaken. Candida albicans was the most common species (47.3%) followed by C. glabrata (25.7%), C. parapsilosis (8.0%), C. tropicalis (6.8%) and Saccharomyces cerevisiae (5.1%). Common predisposing factors were antibacterial therapy (45%), hospitalization in intensive care units (34%), presence of in-dwelling catheters (32%), underlying cancer (23%) and major surgery (11%). Most patients had more than one predisposing factor. Fluconazole alone or in combination with another antifungal agent was the treatment of choice for 86.6% of the cases. Susceptibility testing revealed that 93.5% were susceptible to amphotericin B, 39.6% to itraconazole, 42.8% to fluconazole and 87% to voriconazole. Resistance to azoles was more common among C. glabrata isolates

Chytridiomycosis related mortality in a midwife toad (Alytes obstetricans) in Belgium

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis, contributes to amphibian declines worldwide. Recently, the fungus has shown to be widely distributed in Belgium and the Netherlands, although no clinical cases of the disease have been diagnosed yet. This case report describes the first case of mortality due to chytridiomycosis in Belgium in a wild population of midwife toads (Alytes obstetricans). The presence of clinical chytridiomycosis, combined with the relatively high prevalence of the fungus in Belgium, emphasizes the urgent need for a thorough study on the impact of infection on the native amphibian populations in Belgium

Microsporum gypseum infection in a horse with severe sweet itch

In a horse with severe sweet itch and a history of corticosteroid treatment for this disorder, a single, well-circumscribed and crusty white lesion was present on the thigh. The geophilic dermatophyte species M. gypseum was cultured from the lesion. The lesion was treated topically once a week with an enilconazole emulsion for four weeks. Full recovery was noted after two months. This case suggests that sweet itch combined with systemic corticosteroid treatment, may act as predisposing factor for dermatophyte infection. In addition, the importance of correct isolation and identification of the causal agent is highlighted

In vitro modeling of Batrachochytrium dendrobatidis infection of the amphibian skin

The largest current disease-induced loss of vertebrate biodiversity is due to chytridiomycosis and despite the increasing understanding of the pathogenesis, knowledge unravelling the early host-pathogen interactions remains limited. Batrachochytrium dendrobatidis (Bd) zoospores attach to and invade the amphibian epidermis, with subsequent invasive growth in the host skin. Availability of an in vitro assay would facilitate in depth study of this interaction while reducing the number of experimental animals needed. We describe a fluorescent cell-based in vitro infection model that reproduces host-Bd interactions. Using primary keratinocytes from Litoria caerulea and the epithelial cell line A6 from Xenopus laevis, we reproduced different stages of host cell infection and intracellular growth of Bd, resulting in host cell death, a key event in chytridiomycosis. The presented in vitro models may facilitate future mechanistic studies of host susceptibility and pathogen virulence

Instant killing of pathogenic chytrid fungi by disposable nitrile gloves prevents disease transmission between amphibians

To prevent transmission of the pathogenic chytrid fungi Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal), hygiene protocols prescribe the single use of disposable gloves for handling amphibians. We discovered that rinse water from nitrile gloves instantly kills 99% of Bd and Bsal zoospores. Transmission experiments using midwife toads (Alytes obstetricans) and Bd, and Alpine newts (Ichthyosaura alpestris) and Bsal, show that the use of the same pair of gloves for 2 subsequent individuals does not result in significant transmission of any chytrid fungus. In contrast, handling infected amphibians bare-handed caused transmission of Bsal in 4 out of 10 replicates, but did not result in transmission of Bd. Based on the manufacturer’s information, high resolution mass spectrometry (HRMS) and colorimetric tests, calcium lactate and calcium nitrate were identified as compounds with antifungal activity against both Bd and Bsal. These findings corroborate the importance of wearing gloves as an important sanitary measure in amphibian disease prevention. If the highly recommended single use of gloves is not possible, handling multiple post-metamorphic amphibians with the same pair of nitrile gloves should still be preferred above bare-handed manipulation

Voriconazole, a safe alternative for treating infections caused by the Chrysosporium anamorph of Nannizziopsis vriesii in bearded dragons (Pogona vitticeps)

Dermal and systemic infections caused by the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) are highly prevalent in reptiles and may result in severe disease and high mortality. Due to the high incidence of therapeutic failures, optimizing treatment is required. We first determined in this study the minimal inhibitory concentrations (MIC) of itraconazole, voriconazole, amphotericin B and terbinafine against 32 CANV isolates. For voriconazole, amphotericin B and terbinafine a monomodal MIC distribution was seen, whereas a bimodal MIC distribution was present for itraconazole, indicating acquired resistance in one isolate. Fourteen naturally-infected bearded dragons (Pogona vitticeps), from the same owner, were treated orally with either itraconazole (5 mg/kg q24h) or voriconazole (10 mg/kg q24h). The clinical condition, drug plasma concentrations and the presence of CANV in skin samples were followed. The animals were treated until complete clearance of the fungus. The plasma concentrations of voriconazole and itraconazole exceeded the minimal inhibitory concentrations of the CANV isolates. Elimination of CANV was achieved on average after 27 and 47 days of treatment with itraconazole and voriconazole, respectively. Whereas only 2 out of 7 survived after itraconazole treatment, only a single animal died in the voriconazole treated group. In conclusion, based on a limited number of animals, voriconazole applied at a regimen of 10 mg/kg bodyweight (BW) q24h seems to be a safe and effective antimycotic drug to eliminate CANV infections in bearded dragons