Bol. Asoc. Herpetol. Esp. (2020) 31(2) Preprint-151 Amphibian micro-hotspot at the Mindelo Ornithological Reserve (Porto, Portugal) Guillermo Velo-Antón CIBIO/InBIO, Centro de Investigacão em Biodiversidade e Recursos Genéticos. Instituto de Ciências Agrárias de Vairão 7. Universidade do Porto. Vairão. Portugal. C.e.: [email protected] Fecha de aceptación: 13 de septiembre de 2020. Key words: conservation, diversity, sanctuary. RESUMEN: La reserva ornitológica de Mindelo (Vila do Conde, distrito de Oporto), fue creada en 1957, y es pionera en la conservación de la naturaleza en Portugal. Esto permitió proteger esta pequeña área (380 ha) de la continua degradación del ecosistema costero ocasionada por diversas presiones antrópicas durante las últimas décadas. Su proximidad al mar y diversidad de paisajes proporcionan un clima y hábitat propicios para muchas especies de aves, pero también de anfibios, reptiles y otros grupos, constituyendo un micro refugio para la biodiversidad. En esta nota se proporciona un listado de las especies de anfibios observados durante la pasada década (2011-2020), y una valoración cualitativa de la abundancia de las especies encontradas. Durante este tiempo se ha observado un total de 12 anfibios, representando el 63% de los anfi- bios portugueses, destacando la importancia de este micro refugio para muchos de los anfibios ibéricos y la fragilidad de un espacio que necesita ser gestionado con medidas que garanticen la supervivencia de estas especies. The Mindelo Ornithological Reserve thermal and moisture conditions resulted (MOR) lays in a coastal area of ca. 380 ha. from the influence of the Atlantic Ocean. Its located at the municipality of Vila de Conde heterogeneous landscape, mostly dominated in North Portugal (Figure 1). Its importance by sand dunes, pine forest, and shrubs, with for migratory birds provided the foundations two small coastal lagoons (maintained with to create the first Portuguese area for nature water coming from the two streams crossing conservation in 1957. MOR acts as a refuge this area), and the formation of numerous for biodiversity conservation because it main- temporary ponds and puddles across the re- tains patches of the natural landscape that serve, contribute to the reproduction and once dominated the coastal area in northern survival of amphibian species. All amphibians Portugal, which was continuously degraded inhabiting MOR are water dependent, but and fragmented over the last decades (from show marked differences in life-history traits the 70´s onwards) due to human activities (e.g. oviparous and larviparous reproduction, (e.g. industry, urbanization, agriculture), also sand borrowers and woodland species). resulting in a reduction of wetlands. Thus, Researchers from CIBIO have recently MOR is currently considered an important showed interest in studying some of the am- sanctuary for wildlife. phibians currently present in MOR. In par- This reserve provides optimal climatic ticular, the fire salamander (Salamandra sala- conditions for many ectotherm species, and mandra Linnaeus, 1758), which was used as in particular for amphibians, with buffered the reference population of the subspecies S. 152-Preprint Bol. Asoc. Herpetol. Esp. (2020) 31(2) Foto Albert Martínez-Silvestre Figure 1: Map containing the Mindelo Ornithological Reserve surrounded by urban, agricultural, industrial areas and the Atlantic Ocean. The black dot in the inset represents the location of this reserve in the municipality of Vila de Conde (North Portugal). Figura 1: Mapa de la Reserva Ornitológica de Mindelo rodeada por áreas urbanas, agrícolas e industriales y el Océano Atlántico. El punto negro representa la posición de esta reserva en el municipio de Vila do Conde (norte de Portugal). s. gallaica for genetic (Mulder et al., 2016) and traditional and eDNA methods, however, morphological (Alarcón et al., 2020) studies. In the short-term (over a 10-day period) surveys a recent study, Peixoto et al. (2020) surveyed conducted for this particular study (March some of the puddles and ponds across this 2018) might not represent the full amphibian reserve to evaluate several field and labora- community. Here, I list the number of species tory protocols for amphibian detection using observed over sporadic visits to this reserve environmental DNA (eDNA) methods from since 2011, also giving a qualitative assess- water samples. MOR amphibian community, ment of each species´ abundance. These visits and fire salamanders in particular, served as mostly occurred during rainy nights in the an excellent study system because of the nu- highest peak of activity and the reproductive merous temporary puddles occurring within period of the amphibians present in this re- the reserve, where amphibians can be easily gion (October-November and March-May). I found and detected with traditional sampling found a total of 12 species (eight anurans and methods, allowing for multiple replicates to four urodeles), representing 63% of the Por- evaluate eDNA capture methods and mole- tuguese amphibians (Table 1; Figure 2). This cular approaches. Peixoto et al. (2020) listed constitutes, to the best of my knowledge, the the number of species detected with both highest recorded number of amphibians in Bol. Asoc. Herpetol. Esp. (2020) 31(2) Preprint-153 Portugal. The most abundant species, which cultripes) show high densities during nights can be found in dozens in one single night, with high levels of humidity and mild tem- are the natterjack toad (Epidalea calamita), peratures, as it often occurs across their dis- and the Iberian painted frog (Discoglossus gal- tributions (Recuero, 2014; Velo-Antón & Buckley ganoi). The fire salamander (Salamandra sala- 2015). Three newt species inhabit this reserve: mandra) and the western spadefoot (Pelobates the palmate newt (Lissotriton helveticus), the Figure 2: From top-left to bottom-right the 12 amphibians observed in the Mindelo Ornithological Reserve: Epi- dalea calamita, Discoglossus galganoi, Pelobates cultripes, Lissotriton helveticus, Triturus marmoratus, Salamandra sala- mandra, Lissotriton boscai, Pelodytes atlanticus, Alytes obstetricans, Bufo spinosus, Hyla molleri, Pelophylax perezi. Stroke color of each figure denote the abundance: high (green) and low abundance (blue), and rare (red). Figura 2: De arriba-izquierda a abajo-derecha, las 12 especies de anfibios observados en la Reserva Ornitológica de Mindelo: Epidalea calamita, Discoglossus galganoi, Pelobates cultripes, Lissotriton helveticus, Triturus marmoratus, Salamandra salaman- dra, Lissotriton boscai, Pelodytes atlanticus, Alytes obstetricans, Bufo spinosus, Hyla molleri, Pelophylax perezi. El color del trazado de cada panel denota: abundancia alta (verde), abundancia baja (azul) y especies raras (rojo). 154-Preprint Bol. Asoc. Herpetol. Esp. (2020) 31(2) Table 1: List of the amphibians present in Portugal, To complete their life cycle, amphibians identifying the species observed in the Mindelo Or- present in MOR rely on the numerous sha- nithological Reserve (*), and the relative abundance llow puddles formed across the reserve during of each species. Question marks denote species that might be extinct in the study area. rainy periods, which can become relative lar- Tabla 1: Lista de anfibios presentes en Portugal, ge with episodes of heavy rain or continued identificando las especies observadas en la Reserva rainy days. With the exception of P. cultri- Ornitológica de Mindelo (*), y la abundancia rela- tiva de cada especie. Los signos de interrogación de- pes, which reproduces in stagnant water bo- notan especies que podrían estar extintas en el área dies that can sustain their feeding habits and de estudio. larger tadpole size, all remaining anurans Amphibians Amphibians Abundance and the four urodeles largely frequently use (Portugal) (MOR) (MOR) small puddles for reproduction, laying their Alytes cisternasii low eggs (or larvae in S. salamandra) into these Alytes obstetricans * rare temporary and unpredictable water bodies. Bufo spinosus * Chioglossa lusitanica Salamandra salamandra, P. cultripes, E. cala- high Discoglossus galganoi * high mita and D. galganoi have an apparently wi- Epidalea calamita * Hyla meridionalis der range and use of habitat types compared rare Hyla molleri * to the remaining amphibians. For instance, I low Lissotriton boscai * high often found adults of S. salamandra across the Lissotriton helveticus * high Pelobates cultripes * woodland but also covered in grains in sandy Pelodytes ibericus areas, which indicates the presence of nearby low Pelodytes atlanticus * rare shelters in these particular habitats. Future Pelophylax perezi * Pleurodeles waltl ? efforts should be oriented to spatially charac- Rana iberica ? terize the distribution of each species, as well high Salamandra salamandra * high as to estimate their habitat use, population Triturus marmoratus * Triturus pygmaeus densities and species interactions. Despite this is a protected area, MOR also contains eucalypts and small agricultural pat- marbled newt (Triturus marmoratus), and the ches, allowing the entrance of vehicles used Bosca´s newt (Lissotriton boscai), being the for agricultural activities. I occasionally ob- latter the least abundant of the three. The served roadkilled carcasses of several species Lusitanian parsley frog (Pelodytes atlanticus), (e.g. S. salamandra, D. galganoi, E. calamita, and the common midwife toad (Alytes obs- P. cultripes), likely caused by the use of these tetricans) show low