Geographical patterns of the species richness of helminth parasites of moles (Talpa spp.) in Spain: Separating the effects of sampling effort from those of other conditioning factors

We analysed the viscera of 534 moles (Ta l p a spp.) from 30 of the 47 provinces of peninsular Spain, including 255 individuals of T. europaea from eight provinces, 154 individuals of T. occidentalis from 20 provinces, and 125 unidentified Ta l p a individuals from two provinces. We identified their helminth parasites and determined parasite species richness. We related parasite species richness with sampling effort using both a linear and a logarithmic function. We then performed stepwise linear regressions to predict mole parasite species richness from a small set of selected predictor variables that included sampling effort. We applied the resulting models to forecast T. euro p a e a, T. occidentalis, and Ta l p a spp. parasite species richness in all provinces with recorded host presence, assuming different levels of sampling eff o r t . F i n a l l y, we used partial regression analysis to partition the variation explained by each of the selected variables in the models. We found that mole parasite species richness is strongly conditioned by sampling effort, but that other factors such as cropland area and environmental disturbance have significant independent effects.

DETECTION OF SPORES AND HYPHAE OF ARTWORKS’ BIODETERIOGENIC FILAMENTOUS FUNGI BY RNA-FISH

Filamentous fungi are a threat to the conservation of Cultural Heritage. Thus, detection and identification of viable filamentous fungi are crucial for applying adequate Safeguard measures. RNA-FISH protocols have been previously applied with this aim in Cultural Heritage samples. However, only hyphae detection was reported in the literature, even if spores and conidia are not only a potential risk to Cultural Heritage but can also be harmful for the health of visitors, curators and restorers. Thus, the aim of this work was to evaluate various permeabilizing strategies for their application in the detection of spores/conidia and hyphae of artworks’ biodeteriogenic filamentous fungi by RNA-FISH. Besides of this, the influence of cell aging on the success of the technique and on the development of fungal autofluorescence (that could hamper the RNA-FISH signal detection) were also investigated. Five common biodeteriogenic filamentous fungi species isolated from biodegradated artworks were used as biological model: Aspergillus niger, Cladosporium sp, Fusarium sp, Penicillium sp. and Exophialia sp. Fungal autofluorescence was only detected in cells harvested from Fusarium sp, and Exophialia sp. old cultures, being aging-dependent. However, it was weak enough to allow autofluorescence/RNA-FISH signals distinction. Thus, autofluorescence was not a limitation for the application of RNA-FISH for detection of the taxa investigated. All the permeabilization strategies tested allowed to detect fungal cells from young cultures by RNA-FISH. However, only the combination of paraformaldehyde with Triton X-100 allowed the detection of conidia/spores and hyphae of old filamentous fungi. All the permeabilization strategies failed in the Aspergillus niger conidia/spores staining, which are known to be particularly difficult to permeabilize. But, even in spite of this, the application of this permeabilization method increased the analytical potential of RNA FISH in Cultural Heritage biodeterioration. Whereas much work is required to validate this RNA-FISH approach for its application in real samples from Cultural Heritage it could represent an important advance for the detection, not only of hyphae but also of spores and conidia of various filamentous fungi taxa by RNA-FISH.