Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari: Ixodidae) ticks: prevalence and methods.

Entomopathogenic fungi are commonly found in forested soils that provide tick habitat, and many species are pathogenic to Ixodes scapularis Say, the blacklegged tick. As a first step to developing effective biocontrol strategies, the objective of this study was to determine the best methods to isolate entomopathogenic fungal species from field-collected samples of soils and ticks from an Eastern deciduous forest where I. scapularis is common. Several methods were assessed: (1) soils, leaf litter, and ticks were plated on two types of media; (2) soils were assayed for entomopathogenic fungi using the Galleria bait method; (3) DNA from internal transcribed spacer (ITS) regions of the nuclear ribosomal repeat was extracted from pure cultures obtained from soils, Galleria, and ticks and was amplified and sequenced; and (4) DNA was extracted directly from ticks, amplified, and sequenced. We conclude that (1) ticks encounter potentially entomopathogenic fungi more often in soil than in leaf litter, (2) many species of potentially entomopathogenic fungi found in the soil can readily be cultured, (3) the Galleria bait method is a sufficiently efficient method for isolation of these fungi from soils, and (4) although DNA extraction from ticks was not possible in this study because of small sample size, DNA extraction from fungi isolated from soils and from ticks was successful and provided clean sequences in 100 and 73% of samples, respectively. A combination of the above methods is clearly necessary for optimal characterization of entomopathogenic fungi associated with ticks in the environment.

Ectomycorrhizal responses to organic and inorganic nitrogen sources when associating with two host species.

While it is established that increasing atmospheric inorganic nitrogen (N) deposition reduces ectomycorrhizal fungal biomass and shifts the relative abundances of fungal species, little is known about effects of organic N deposition. The effects of organic and inorganic N deposition on ectomycorrhizal fungi may differ because responses to inorganic N deposition may reflect C-limitation. To compare the effects of organic and inorganic N additions on ectomycorrhizal fungi, and to assess whether host species may influence the response of ectomycorrhizal fungi to N additions, we conducted an N addition experiment at a field site in the New Jersey pine barrens. Seedlings of two host species, Quercus velutina (black oak) and Pinus rigida (pitch pine), were planted at the base of randomly-selected mature pitch pine trees. Nitrogen was added as glutamic acid, ammonium, or nitrate at a rate equivalent to 227.5 kg ha(-1) y(-1) for eight weeks, to achieve a total application of 35 kg ha(-1) during the 10-week study period. Organic and inorganic N additions differed in their effects on total ectomycorrhizal root tip abundance across hosts, and these effects differed for individual morphotypes between oak and pine seedlings. Mycorrhizal root tip abundance across hosts was 90 % higher on seedlings receiving organic N compared to seedlings in the control treatment, while abundances were similar among seedlings receiving the inorganic N treatments and seedlings in the control. On oak, 33-83 % of the most-common morphotypes exhibited increased root tip abundances in response to the three forms of N, relative to the control. On pine, 33-66 % of the most-common morphotypes exhibited decreased root tip abundance in response to inorganic N, while responses to organic N were mixed. Plant chemistry and regression analyses suggested that, on oak seedlings, mycorrhizal colonization increased in response to N limitation. In contrast, pine root and shoot N and C contents did not vary in response to any form of N added, and mycorrhizal root tip abundance was not associated with seedling N or C status, indicating that pine received sufficient N. These results suggest that in situ organic and inorganic N additions differentially affect ectomycorrhizal root tip abundance and that ectomycorrhizal fungal responses to N addition may be mediated by host tree species.

Drug-induced hypersensitivity reaction: A case of simultaneous thyroiditis and fulminant type 1 diabetes.

A 43-year-old incarcerated man with AIDS was hospitalized for 30 pounds weight loss and diffuse pruritic rash. Three months prior, he was started on dapsone for Pneumocystis jiroveci pneumonia prevention. Biochemical evaluation was remarkable for eosinophilia, thrombocytopenia, acute renal insufficiency, transaminitis, thyrotoxicosis, and significant hyperglycemia (450 mg/dl; nl, 65-99). His hemoglobin A1c level was 5.9% (nl, 4.1-5.6). Thyroid-stimulating immunoglobulin, glutamic acid decarboxylase, and islet cell autoantibodies were within the normal range. He was found to have acute interstitial nephritis based on renal biopsy. He was diagnosed with hypersensitivity reaction due to dapsone. The patient was managed with a tapering dose of corticosteroid, beta-blocker, and multiple daily injections of insulin. The symptoms and biochemical disturbances including thyrotoxicosis resolved within a few weeks. Insulin requirements decreased but diabetes did not resolve with hemoglobin A1c of 6.1% a year after hospitalization. To our knowledge, this is the first case of hypersensitivity reaction due to dapsone causing simultaneous fulminant type 1 diabetes and thyroiditis.