The distribution of the chytrid fungus Batrachochytrium salamandrivorans continues to expand in Europe. During 2014–2018, we collected 1,135 samples from salamanders and newts in 6 countries in Europe. We identified 5 cases of B. salamandrivorans in a wild population in Spain but none in central Europe or the Balkan Peninsula.
Environmental DNA (eDNA) is becoming an indispensable tool in biodiversity monitoring, including the monitoring of invasive species and pathogens. Aquatic chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) are major threats to amphibians. However, the use of eDNA for detecting these pathogens has not yet become widespread, due to technological and economic obstacles. Using the enhanced eDNA approach (a simple and cheap sampling protocol) and the universally accepted qPCR assay, we confirmed the presence of Bsal and Bd in previously identified sites in Spain, including four sites that were new for Bsal. The new approach was successfully tested in laboratory conditions using manufactured gene fragments (gBlocks) of the targeted DNA sequence. A comparison of storage methods showed that samples kept in ethanol had the best DNA yield. Our results showed that the number of DNA copies in the Internal Transcribed Spacer region was 120 copies per Bsal cell. Eradication of emerging diseases requires quick and cost-effective solutions. We therefore performed cost-efficiency analyses of standard animal swabbing, a previous eDNA approach, and our own approach. The procedure presented here was evaluated as the most cost-efficient. Our findings will help to disseminate information about efforts to prevent the spread of chytrid fungi.
Batrachochytrium salamandrivorans (Bsal), a pathogenic fungus causing the fatal disease chytridiomycosis in amphibians, was likely introduced to Europe through the trade in pet salamanders from Asia and then escaped into wild populations. Among European countries, Spain has a large number of private breeders and keepers of pet salamanders, and cases of Bsal in wild and captive populations already have been confirmed there. However, surveillance for the pathogen in Spanish collections of amphibians is sparse. Therefore, assisted by private owners and breeders, we surveyed 10 amphibian collections and analysed a total of 317 samples for presence of Bsal. All of our analyses yielded negative results. However, this apparent lack of Bsal cases in captivity should not encourage relaxation of vigilance, quarantine efforts or good practices. Because amphibian collections represent highly dynamic environments (animals are coming in and out), the pathogen could easily be introduced into a collection by new individuals. Any case of Bsal infection in captive animals could lead to its further spread to wild populations of susceptible species, potentially decimating them, and thus should be prevented.
BackgroundBirds are one of the groups involved in the development of Sarcocystis Lankester, 1882, serving either as intermediate or definitive hosts. The white-tailed sea eagle Haliaeetus albicilla (Linnaeus, 1758), red kite Milvus milvus (Linnaeus, 1758) (both Accipitriformes) and common starlings Sturnus vulgaris Linnaeus, 1758 (Passeriformes) were examined to elucidate their participation in the development of Sarcocystis, as well as to determine the specific identity of the parasites based on morphological and especially molecular analyses.MethodsIn 2020–2021, one white-tailed eagle, one red kite and five common starlings were parasitologically examined for the presence of Sarcocystis using flotation centrifugation coprological method and by wet mounts of intestinal mucosa scrapings and/or muscle samples. Positive samples were processed by light microscopy, histologically and followed molecularly at four genetic markers (18S rRNA, 28S rRNA, ITS1 and cox1).ResultsThe white-tailed eagle harboured oocysts/sporocysts of S. arctica Gjerde et Schulze, 2014 in the intestinal mucosa, while the intestinal mucosa of the red kite and breasts and leg muscles of one common starling were positive to S. halieti Gjerde, Vikøren et Hamnes, 2018. Sequences from eagle shared 99.6 − 100% identity with each other and S. arctica in the red fox (V. vulpes Linnaeus, 1758) from the Czech Republic. Sequences from the common starling and red kite shared 100% identity with each other and with S. halieti in the great cormorant (P. carbo [Linnaeus, 1758]) from Lithuania and H. albicilla from Norway.ConclusionsThe white-tailed sea eagle (H. albicilla) acts as natural definitive host of S. arctica, whereas the common starling (St. vulgaris) and red kite (M. milvus) represent intermediate and definitive hosts, respectively, for S. halieti.
Background: Species of Sarcocystis are parasitic protozoan in poikilothermic and homeothermic animals. Out of the 25 valid avian species, none has been reported in birds of the order Musophagiformes, as the great blue turaco Corythaeola cristata (Vieillot, 1816), which is an endemic bird inhabiting Central and Western Africa. The examination of great blue turacos imported from the Central Africa Republic to Czech Republic allowed the morphological and molecular characterization of a new species of Sarcocystis.Methods: Four turacos imported from the Central Africa Republic to private breeder (Czech Republic) were parasitologically examined through wet mounts of breast, heart and legs muscles for the presence of sarcocysts. Found parasites were molecular and histologically studied by 4 loci (18S rRNA, 28S rRNA, ITS1 and cox1) and haematoxylin and eosin stain, respectively. Results: Three out of 4 examined birds (prevalence: 75%) harboured numerous sarcocysts in breast and leg muscles. No macroscopic lesions where observed. Sarcocysts were microscopic, elongate and ribbon-shaped with a wall characterised by the presence of finger-shaped villar protrusions. The new species is molecularly similar to S. albifronsi, S. anasi, S. atraii, S. chloropusae, S. lari and S. rileyi) in the four loci. Conclusions: This is the first species of Sarcocystis in a musophagiform bird around the world. Genetically, S. cristata sp. nov. represents a distinct species. Phylogenetic analyses are useful for predicting potential definitive hosts of the new Sarcocystis species.
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