Co-occurrence of anatoxin-a and microcystins in Lake Garda and other deep subalpine lakes

Cyanotoxins are a global concern in freshwaters and eutrophication and climate changes can have synergistic effects in exacerbating the problem. The deep perialpine lakes are a group of lakes of huge economic and naturalistic importance located at the border of the Alps. At the southern border of the Italian and Swiss Alps, the largest waterbodies include the lakes Garda, Iseo, Como, Lugano and Maggiore (Deep Subalpine Lakes, DSL). Together with eutrophication (during the 1960s and 1970s) and re-oligotrophication (from the 1990s onward) these lakes have been experiencing warming and increase of the water column stability. These changes had a strong impact on the phytoplankton (including cyanobacteria) community. Four DSL (lakes Garda, Iseo, Como and Lugano) have been studied with the aim of comparing their toxic potential. For one of them (Lake Garda) an 8 years survey was conducted, allowing a long-term trend analysis. Toxin analysis was conducted on a monthly basis by targeted LC-MS/MS. A screening for anatoxins, cylindrospermopsins, saxitoxins, microcystins (MCs) and nodularins was carried out. Among all the listed toxins, only one anatoxin and five MCs were detected in the lakes. In particular, the alkaloid anatoxin-a (ATX) was found dominant in lakes Garda, Iseo and Como, and absent in Lake Lugano; the MC-[D-Asp3]RR was found as the most abundant MC in all four lakes. Four other less abundant MCs were also found. The two major toxins are produced by two different cyanobacteria, Tychonema bourrellyi (J.W.G. Lund) Anagnostidis & Komárek and Planktothrix rubescens (De Candolle ex Gomont) Anagnostidis & Komárek, which share however a number of ecological traits. Peaks of these toxins occurred in warmer months (typically between May and September) in the thermocline layer (around 20 m, in the considered lakes). In summer 2016, the highest concentrations of ATX and total MCs were registered in Lake Iseo (1100 and 430 ng L–1, respectively), while in the other lakes values were approximately twice lower. In the lakes where it was present, ATX peak levels were much higher than MCs, thus highlighting the necessity of including ATX in the procedures of risk assessment. The importance of ATX is expected to further grow in the future with respect to MCs, as demonstrated by the long-term trend analysis carried out in Lake Garda that showed a clear decline for MCs from 2009 till 2016 and a relative constancy of ATX.