Fine-scale cryogenic sampling of planktonic microbial communities: Application to toxic cyanobacterial blooms
#CryoSamplers #CyanoResearch @JonathanPuddick Jonathan Puddick, Susie Wood, Eric Goodwin, Ian Hawes, Daniel Dietrich & David Hamilton ‘ed #CyanoResearch
Prof. David Hamilton Prof. Ian Hawes Prof. Daniel Dietrich Dr. Susie Wood Bloom-forming Microcystis Scum-forming Microcystis Microcystins
. Most frequently detected freshwater cyanotoxin . Produced by many cyanobacteria genera . >130 structural variants identified . Potent hepatotoxins . Liver tumour promoters Toxin Production: From the laboratory….. ….and into the field Natural Population Surveys
Blue-Green Algae Finer-Scale Profiling of ‘Tranquility Bay’ Mesocosm Investigations Lander Profiling of Microcystis Scums Mixed System Photosynthesising Scum Respiring Scum
DO (saturation %) DO (saturation %) DO (saturation %) 0 50 100 150 200 250 0 50 100 150 200 250 0 50 100 150 200 250 0 0 0
2 2 2
4 4 4
6 6 6
8 8 8
Depth(mm) Depth(mm) Depth(mm)
10 10 10
12 12 12
14 14 14 pH pH pH 6 8 10 12 6 8 10 12 6 8 10 12 0 0 0
2 2 2
4 4 4
6 6 6
8 8 8
Depth(mm)
Depth(mm) Depth(mm)
10 10 10
12 12 12
14 14 14 Cryo-Sampling of Cyanobacteria Vertical Profiles using ‘Cold Finger’ Fine-scale Vertical Profiles of Toxin Levels Scum Formation
80% MeOH Clarify LC-MS Dual Extraction of Cryo-Samples
Heat at 99 °C LC-MS DNA DNA
Supplement with
0.1% Tween-20 .
.Quantify toxic genotypes .McyE gene PCR .Other communities .Microcystis 16S gene? .Cyanobacterial 16S gene? Microcystin Cell Quota
Scum Formation
of toxin percell) toxin of
pg MC Cell Quota MC ( Cell Quota Isolated Communities using ‘Colony Catcher’
High-Throughput DNA Sequencing
Cyanobacterial ITS Primers High-Throughput Sequencing Microcystis Diversity in Colonies Microcystis Diversity in Lake Water Microcystis Diversity at Lake Rotorua Cryogenic Sampling of Communities Advantages Hindrances Field-based (in situ) Cryogenic Substances Instant Preservation Slightly Tedious Increased Precision Ice Crystal Disruption
Check it out in Limnology and Oceanography Methods or on vimeo.....#CyanoResearch Acknowledgements . Marsden Fund of the Royal Society of NZ (12-UOW-087) . NZ Ministry of Business, Innovation & Employment (UOWX0505) . Royal Society IRSES exchange funding
. Ngāi Tahu, the Kahu Family & the Kaikoura locals
. James Topliss, Richard Topliss, Beth Wood, John Wood, Hugo Borges, Shelley Rogers, Konstanze Steiner, Debin Meng, Caroline Hodges, Oonagh Daly, Josephin Brandes, Marcia Maier, Oliver Baudendistel, Sarah Dorner, Sarah Johns, Laura Biessy, Janet Adamson, Veronica Beuzenberg, Tim Dodgshun, Marc Jary and Aaron Quarterman Email: [email protected] Twitter: @JonathanPuddick Vimeo: #CyanoResearch