Emerging infectious diseases caused by fungal taxa are increasing and are placing a substantial burden on economies and ecosystems worldwide. Of the emerging fungal diseases, chytridomycosis caused by the fungus Batrachochytrium dendrobatidis (hereafter Bd) is linked to global amphibian declines. Amphibians have innate immunity, as well as additional resistance through cutaneous microbial communities. Despite the targeting of bacteria as potential probiotics, the role of fungi in the protection against Bd infection in unknown. We used a four-part approach, including high-throughput sequencing of bacterial and fungal communities, cultivation of fungi, Bd challenge assays, and experimental additions of probiotic to Midwife Toads (Altyes obstetricans), to examine the overlapping roles of bacterial and fungal microbiota in pathogen defense in captive bred poison arrow frogs (Dendrobates sp.). Our results revealed that cutaneous fungal taxa differed from environmental microbiota across three species and a subspecies of Dendrobates spp. frogs. Cultivation of host-associated and environmental fungi realved numerous taxa with the ability to inhibit or facilitate the growth of Bd. The abundance of cutaneous fungi contributed more to Bd defense (~45% of the fungal community), than did bacteria (~10%) and frog species harbored distinct inhibitory communities that were distinct from the environment. Further, we demonstrated that a fungal probiotic therapy did not induce an endocrine-immune reaction, in contrast to bacterial probiotics that stressed amphibian hosts and suppressed antimicrobial peptide responses, limiting their long-term colonization potential. Our results suggest that probiotic strategies against amphibian fungal pathogens should, in addition to bacterial probiotics, focus on host-associated and environmental fungi such as Penicillium and members of the families Chaetomiaceae and Lasiosphaeriaceae.

SummaryReduced carbon (C) assimilation during prolonged drought forces trees to rely on stored C to maintain vital processes like respiration. It has been shown, however, that the use of carbohydrates, a major C storage pool and apparently the main respiratory substrate in plants, strongly declines with decreasing plant hydration. Yet no empirical evidence has been produced to what degree other C storage compounds like lipids and proteins may fuel respiration during drought.We exposed young scots pine trees to C limitation using either drought or shading and assessed respiratory substrate use by monitoring the respiratory quotient, d13 C of respired CO 2 and concentrations of the major storage compounds, that is, carbohydrates, lipids and amino acids. Only shaded trees shifted from carbohydrate-dominated to lipid-dominated respiration and showed progressive carbohydrate depletion. In drought trees, the fraction of carbohydrates used in respiration did not decline but respiration rates were strongly reduced. The lower consumption and potentially allocation from other organs may have caused initial carbohydrate content to remain constant during the experiment.Our results suggest that respiratory substrates other than carbohydrates are used under carbohydrate limitation but not during drought. Thus, respiratory substrate shift cannot provide an efficient means to counterbalance C limitation under natural drought.

Photosynthesis and respiration are major components of the plant carbon balance. During stress, like drought, carbohydrate supply from photosynthesis is reduced and the Krebs cycle respiration must be fueled with other stored carbon compounds. However, the dynamics of storage use are still unknown. The respiratory quotient (RQ, CO2 released per O2 consumed during respiration) is an excellent indicator of the nature of the respiration substrate. In plant science, however, online RQ measurements have been challenging or even impossible so far due to very small gas exchange fluxes during respiration. Here we apply cavity-enhanced multi-gas Raman spectrometry (CERS) for online in situ RQ measurements in drought-tolerant pine (Pinus sylvestris [L.]) and drought-intolerant spruce (Picea abies [L. H. Karst]). Two different treatments, drought and shading, were applied to reduce photosynthesis and force dependency on stored substrates. Changes in respiration rates and RQ values were continuously monitored over periods of several days with low levels of variance. The results show that both species switched from COH-dominated respiration (RQ = 1.0) to a mixture of substrates during shading (RQ = 0.77-0.81), while during drought only pine did so (RQ = 0.75). The gas phase measurements were complemented by concentration measurements of non-structural carbohydrates and lipids. These first results suggest a physiological explanation for greater drought tolerance in pine. CERS was proven as powerful technique for non-consumptive and precise real-time monitoring of respiration rates and respirational quotients for the investigation of plant metabolism under drought stress conditions that are predicted to increase with future climate change.

Rates of nest predation have frequently been shown to differ between fragmented and unfragmented habitats, but have rarely been compared among natural habitats in the same geographic region. In this study, artificial nests of two types (open cup and domed) were placed in four habitats (mangroves, monsoon rainforests, eucalypt woodlands and paperbark swamps) over 12 months in three localities near Darwin in the Australian monsoon tropics to determine the effects of habitat, season and nest type on the rate of nest predation. A quail egg and a similarly coloured plasticine egg were placed in each nest. Habitat had a strong effect on nest predation rates, with nests in mangroves experiencing predation rates more than four times higher than those in eucalypt woodlands and paperbark swamps. Despite the strong rainfall seasonality of the region, there was no consistent seasonal variation in nest predation rates. Nest type also had little influence on predation rates, except in paperbark swamps where open cup nests suffered a higher predation rate than domed nests. The study indicates that generalised nest predation rates for tropical regions, even for small areas (e.g. <17 km radius), might overlook substantial variation between habitats. Such variation confounds purported differences in nest predation rates between tropical and temperate regions.

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