Acute Toxicity of Carbon Nanotubes, Carbon Nanodots, and Cell-Penetrating Peptides to Freshwater Cyanobacteria

Synthetic non-metallic nanoparticles (NMNPs) such as carbon nanotubes (CNTs), carbon nanodots (CNDs), and cell-penetrating peptides (CPPs) have been explored to treat harmful algal blooms. However, their strain-specific algicidal activities have been rarely investigated. Here we determined their acute toxicity to nine freshwater cyanobacterial strains belonging to seven genera, including <i>Microcystis aeruginosa</i> UTEX 2386, <i>M. aeruginosa</i> UTEX 2385, <i>M. aeruginosa</i> LE3, <i>Anabaena cylindrica</i> PCC 7122, <i>Aphanizomenon</i> sp. NZ, <i>Planktothrix agardhii</i> SB 1810, <i>Synechocystis</i> sp. PCC 6803, <i>Lyngbya</i> sp. CCAP 1446/10, and <i>Microcoleus autumnale</i> CAWBG635 ATX. We prepared in-house three batches of CNDs using glucose (CND-G) or chloroform and methanol (CND-C/M) as the substrate and one batch of single-walled CNTs (SWCNTs). We also ordered a commercially synthesized CPP called γ-Zein-CADY. The axenic laboratory culture of each cyanobacterial strain was exposed to an NMNP at two dosage levels (high and low, with high = 2 × low) for 48 h, followed by measurement of five endpoints. The endpoints were optical density (OD) at 680 nm (OD₆₈₀) for chlorophyll-a estimation, OD at 750 nm (OD₇₅₀) for cell density, instantaneous pigment fluorescence emission (FE) after being excited with 450 nm blue light (FE₄₅₀) for chlorophyll-a or 620 nm red light (FE₆₂₀) for phycocyanin, and quantum yield (QY) for photosynthesis efficiency of photosystem II. The results indicate that the acute toxicity was strain-, NMNP type-, dosage-, and endpoint-dependent. The two benthic strains <i>Microcoleus autumnale</i> and <i>Lyngbya</i> sp. were more resistant to NMNP treatment than the other seven free-floating strains. SWCNTs and fraction A14 of CND-G were more toxic than CND-G and CND-C/M. The CPP was the least toxic. The high dose generally caused more severe impairment than the low dose. OD₇₅₀ and OD₆₈₀ were more sensitive than FE₄₅₀ and FE₆₂₀. QY was the least sensitive endpoint. The strain dependence of toxicity suggested the potential application of these NMNPs as a target-specific tool for mitigating harmful cyanobacterial blooms.