Know Your Enemy: Cellana's Successful Strategy for Dealing with Contaminants in Algal Mass Culture
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1 Know Your Enemy: Cellana's Successful Strategy for Dealing with Contaminants in Algal Mass Culture Charles J. O Kelly Director, Research 2 October 2014 Page 1 Cellana 2014 Cellana Inc. 2014
2 Algae Are Crop Plants Crops Need Protection From Pests / Disease Page 2 Cellana 2014
3 Investing In Crop Protection Identification (How accurately? How quickly?) Behavior Prey range Virulence Life cycle Transmission Vector(s) Susceptibilities Page 3 Cellana 2014
4 Investing In Crop Protection Identification (How accurately? How quickly?) Behavior Prey range Virulence Life cycle Transmission Vector(s) Susceptibilities European corn borer, Ostrinia nubilalis (larva) Page 4 Cellana 2014
5 The State of Algal Crop Protection Identification difficult Most identifications crude ( ciliates / rotifers / amoebae / foreign algae ) Guidebooks nonexistent Carney & Lane 2014 review: 16 taxa of algivorous protozoa named only 9 of the 16 named to species only 3 of the 16 studied in algal mass culture systems 2 of the algal mass culture species new to science 10 µm Behavior and Transmission poorly understood Paraphysoderma sedebokerense, a chytrid fungus, parasitizing Haematococcus. Described Image: Hoffman Y et al., Mycological Research 112: 70, Page 5 Cellana 2014
6 Crop Protection Case Study 1. Identification Page 6 Page 6 Cellana 2014 Cellana 2014
7 Who s eating the profits - really? Culture studies on 5 cell types with ingested algae No growth on target alga: one amoeba, two ciliates Slow growth on target alga: one amoeba Rapid growth on target alga: one amoeba Consistent with observations in large-scale production Page 7 Cellana 2014
8 Who s eating the profits - really? DNA sequence studies assign amoeba to species Neoparamoeba branchiphila Described in 2005 Associated with gills of salmonid fishes affected with amoebic gill disease (not the cause of the disease) No prior record of algivory for any N. species Page 8 Cellana 2014
9 Crop Protection Case Study 2A. Behavior Prey Range Page 9 Page 9 Cellana 2014 Cellana 2014
10 Neoparamoeba: gourmet or gourmand? Susceptible algae Diatoms Some Haptophytes Resistant algae Some haptophytes Green algae Eustigmatophytes Dinoflagellates Cryptophytes Page 10 Cellana 2014
11 Neoparamoeba: gourmet or gourmand? Grows equally well, or better, on bacteria vs. algae Not all bacteria support growth Variations between strains in grazing on both algae and bacteria Marinobacter sp. 1 Marinobacter sp. 2 Halomonas sp. Nitratireductor sp. 1 Labrenzia aggregata Roseivivax sp. Labrenzia sp. Marinobacter hydrocarbonoclasticus Sulfitobacter pontiacus Nb O3a Nb R1 Nb K3 Nitratireductor sp. 2 Page 11 Cellana 2014
12 Crop Protection Case Study 2B. Behavior Virulence Page 12 Page 12 Cellana 2014 Cellana 2014
13 Components of virulence Grazing rate 1-3 prey cells grazer -1 h -1 Specific growth rate Considerably more variability in specific growth rate among grazers than in grazing rate Grazing of Neoparamoeba (03a, R1, KPF3), Paramoeba (08pp), and Thecamoeba (8t) amoebae on the diatom Skeletonema marinoi, incubated in the dark at 23 C. Control: no amoebae. Page 13 Cellana 2014
14 The significance of specific growth rate Appearance of organisms in batch and semi-continuous culture a function of specific growth rate (initial inoculum a relatively small contributor) Model assumes no loss of cells Graph at right shows progression of organism with specific growth rate of 2 d -1 and initial inoculum of cells L -1 Forehead H, O Kelly CJ, Bioresource Technology 129: 329, 2013 Page 14 Cellana 2014
15 The significance of specific growth rate Neoparamoeba branchiphila on favored prey has specific growth rate ca. 2 d -1 Timing of appearance of N. branchiphila in large-scale production closely followed model predictions Dashed line shows limit of detection of contaminants via microscopic monitoring. Forehead H, O Kelly CJ, Bioresource Technology 129: 329, 2013 Page 15 Cellana 2014
16 Crop Protection Case Study 2C. Behavior Life Cycle Page 16 Page 16 Cellana 2014 Cellana 2014
17 WYSIWYG A one-phase life cycle: trophic cells (amoebae) No cysts No resting stages No obvious sexual reproduction Control the trophs, control the pest Page 17 Cellana 2014
18 Crop Protection Case Study 3. Transmission Vector and Susceptibilities Page 18 Page 18 Cellana 2014 Cellana 2014
19 Airborne or Waterborne? Airborne transmission of Neoparamoeba? No cysts No desiccation-resistant stages No evidence for amoebae in air column samples Waterborne transmission of Neoparamoeba? Ability of amoebae to subsist on bacteria suggests possibility of reservoirs Must survive UV, several degrees of water filtration Page 19 Cellana 2014
20 Waterborne! Up to 12 Neoparamoeba cells per 100 L detected in filtered seawater used in production Only 1 cell per 1000 L needed for contamination in 14 days per model Neoparamoeba cells survived 300 mj UV in laboratory tests Production UV was delivering ca. 150 mj Page 20 Cellana 2014
21 Actions Page 21 Page 21 Cellana 2014 Cellana 2014
22 ALDUO Closed System Photobioreactors (PBRs) + Open System Open Raceway Ponds Clean water Clean procedures Resistant strains Clean water Clean inoculum Pond cycle management Page 22 Cellana 2014
23 ALDUO Closed System Photobioreactors (PBRs) + Open System Open Raceway Ponds Clean water Clean procedures Resistant strains Clean water Clean inoculum Pond cycle management Page 23 Cellana 2014
24 ALDUO Closed System Photobioreactors (PBRs) + Open System Open Raceway Ponds Clean water Clean procedures Resistant strains Clean water Clean inoculum Pond cycle management Page 24 Cellana 2014
25 Thanks to: Geneva Mottet Dan Burton Yana Eglit Page 25 Page 25 Cellana 2014 Cellana 2014
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