PIC-LCP-15.19 Anabaena sp. Capitão. Oct/2015. SMG-LES-16.20 Nostoc sp. Empadadas Sul. Mar/2016. SMG-LST-16.21 Aphanizomenon gracile San6ago.
Isola&on, produc&on and maintenance of axenic cultures of freshwater species of Cyanobacteria from the Azorean Islands Rúben Luz1, Rita Cordeiro1, Amélia Fonseca1 & Vítor Gonçalves1,2 1 Departamento de Biologia da Universidade dos Açores, Ponta Delgada, Açores, Portugal 2 CIBIO, Centro de InvesIgação em Biodiversidade e Recursos GenéIcos, InBIO Laboratório Associado, Ponta Delgada, Portugal
IntroducIon The Azorean islands are rich in freshwaters, such as springs, streams, wetlands and lakes, which are hotspots of biodiversity1. Cyanobacteria are amongst the most diverse group of microorganisms that inhabit these ecosystems2. Especially in lakes, as result of eutrophicaIon, cyanobacteria have become the dominant group. Due to their wide diversity of secondary metabolites, where some are toxic (cyanotoxins) but others are believed to have possible use as biocide, anIfungal, anIbioIc and anIcancer substances, the isolaIon and culture of cyanobacteria for their biochemical and geneIc characterizaIon are of great interest2. The aim of this study is to create a culture bank of Azorean cyanobacteria strains, that allows geneIc characterizaIon and bioprospecIng of cyanobacteria from the Azorean Islands.
Methods Samples from 24 lakes in São Miguel, Pico, Flores and Corvo islands (Azores Archipelago) are being collected (Fig. 1) seasonally (winter, spring, summer and autumn) since 2015 and the cyanobacteria present isolated and maintained in axenic cultures. Several technics were used for isolaIon, such as diluIon (Fig. 2) and inverted microscope direct pipeong (Fig. 3). Cultures were maintained in three different mediums (BG-11, BG-11(N) and TFI+M) under controlled temperature and light condiIons3 (Fig. 4). Standard isolaIon and culture techniques were adapted according to the characterisIcs of the isolated strains or the desired taxa to isolate.
Figure 1 – Sampling with net
Figure 2 – Samples diluted in 100 mL Erlenmeyers
Figure 4 – Growing and maintenance chambers
Figure 3 – Inverted microscope in flow chamber
Table 1 – List of cyanobacteria isolates in the bank
Figure 5 – Planktolyngbya sp., SMG-LFR-14.1
Figure 8 – Oscillatoria tenuis, SMGLAR-15.13
Figure 6 – Aphanothece sp., SMG-LC-14.5
Figure 7 – Microcys>s flos-aquae, SMGLFR-14.6
Figure 9 - Nostoc sp., SMGLES-16.20
Figure 10 - Aphanizomenon gracile, SMG-LST-16.21
A
B
Isolate Taxa code
Lake
Date of collecIon
SMG-LFR-14.1
Planktolyngbya sp.
Furnas
Oct/2014
SMG-LA-14.2
Pseudanabaena sp.
Azul
Oct/2014
SMG-LRSD-14.3 Pseudanabaena sp.
Rasa da Serra Devassa Oct/2014
SMG-LRST-14.4 Pseudanabaena sp.
Rasa das Sete Cidades Oct/2014
SMG-LCA-14.5 Aphanothece sp.
Carvão
Oct/2014
SMG-LFR-14.6
Microcys>s flos-aquae Furnas
Nov/2014
SMG-LST-15.7
Pseudanabaena sp.
Mar/2015
SMG-LFR-15.8
Microcys>s flos-aquae Furnas
Apr/2015
SMG-LFR-15.9
Microcys>s aeruginosa Furnas
Apr/2015
SMG-LES-15.10 Oscillatoria tenuis
Empadadas Sul
SMG-LRSC-15.11 Oscillatoria tenuis
Rasa das Sete Cidades July/2015
SMG-LRSD-15.12 Pseudanabaena sp.
Rasa da Serra Devassa July/2015
SMG-LAR-15.13 Oscillatoria tenuis
Areeiro
July/2015
SMG-LAR-15.14 Pseudanabaena sp.
Areeiro
July/2015
SMG-LSB-15.15 Pseudanabaena sp.
São Brás
July/2015
SMG-LPR-15.16 Oscillatoria tenuis
Pico d’el Rei
July/2015
SMG-LCD-15.17 Pseudanabaena sp.
Caiado
Oct/2015
FLO-LN-15.18
Anabaena sp.
Negra
Oct/2015
PIC-LCP-15.19
Anabaena sp.
Capitão
Oct/2015
Empadadas Sul
Mar/2016
SMG-LES-16.20 Nostoc sp.
Figure 11 – Anabaena planctonica, SMGLES-16.22
Figure 12 – Calothrix sp., SMGLF-16.23
Figure 13 – UnidenIfied Nostocales. A: Azul lake; B: São Brás lake
At the moment we have successfully isolated and produced 25 axenic cultures of cyanobacteria strains belonging to several genera: Planktolyngbya (1 strain); Pseudanabaena (8 strains); Aphanothece (1 strain); Microcys>s (3 strains); Oscillatoria (4 strains); Anabaena (3 strains); Nostoc (3 strains); Aphanizomenon (1 strain); Calothrix (1 strain) (Table 1 and Figures 5-13).
SMG-LST-16.21 Aphanizomenon gracile SanIago
Mar/2016
SMG-LES-16.22 Anabaena planctonica Empadadas Sul
Mar/2016
SMG-LFR-16.23 Calothrix sp,
Furnas
Mar/2016
PIC-LPX-16.24
Peixe
Mar/2016
São Brás
Mar/2016
Nostoc sp.
SMG-LSB-16.25 Nostoc sp.
This bank will allow the first studies ever made in Azores with pure cultures of cyanobacteria, giving the chance to study these species as sources for new valuable products and applicaIons.
References
May/2015
Final remark
Results
Bank Uses & Benefits
SanIago
e.g. toxins
GeneIc and chemical studies
BioprosprecIon
Secondary metabolites
1. Strayer, D. L. & D. Dudgeon (2010). Freshwater biodiversity conservaIon: recent progress and future challenges. Journal of the North American Benthological Society 29(1):344-358. 2. Whiaon, B. A. (Ed.). (2012). Ecology of cyanobacteria II: their diversity in space and >me. Springer Science & Business Media. 3. Rippka, R. (1988). IsolaIon and purificaIon of cyanobacteria. Methods in Enzymology 167:3-27.
IdenIficaIon Health prevenIon Biological acIviIes Biotechnology
