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or Extreme Halophiles: Host-Virus Dynamics

Isolation and Physiological Characterization of a Halophilic Host-Phage System from Solar Salterns in Baja California, Mexico. or Extreme Halophiles: Host-Virus Dynamics. Hypersaline Environments.

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or Extreme Halophiles: Host-Virus Dynamics

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  1. Isolation and Physiological Characterization of a Halophilic Host-Phage System from Solar Salterns in Baja California, Mexico orExtreme Halophiles: Host-Virus Dynamics

  2. Hypersaline Environments • Hypersaline >100 g/liter of total salt; extreme hypersaline > 250 g/liter (up to saturation level of salt) • Thalassohaline or athalassohaline (marine composition vs. non-marine composition) • Great Salt Lake, Dead Sea, solar salterns (ESSA), hypersaline alkaline lakes (East Africa, China) • Abundant microbial life: • Archaeal - require very high [salt] to survive • Bacterial - tolerate wide range of salinities (low to high) • Eucaryal - (Dunaliella: red & green; Picocystis; fungi)

  3. Questions about Halophilic Viruses • What is the diversity of hosts and their phages? • What is ecological range of viruses? Host specificities? • How do halophilic phages affect hosts? • Bacterial community structure (host mortality) • Gene transfer • Are there specific adaptations in halophage to survive in extreme hypersaline environments? • Hosts utilize more acidic amino acid residues and more salt bridges in proteins • fCH1, haloalkaliphilic virus  IEF showed acidic capsid proteins

  4. Field Site: Exportadora de Sal, Guerrero Negro, Baja California, Sur, Mexico

  5. Field Site: Exportadora de Sal, Guerrero Negro, Baja California, Sur, Mexico Guerrero Negro Pond #9…?

  6. Study Site: Exportadora de Sal (ESSA), Baja California, Sur, Mexico • Solar saltern located in Guerrero Negro Lagoon • Salinity from seawater to saturation (~48 g/l to >300 g/l) • Water temperature: 16.2oC – 19oC Pond #9, gypsum crystals Pond #9 on right side Pond #11 Bitterns pond Crystallization pond Crystallizer Channel

  7. Water Sampling Protocol • Visited extreme hypersaline ponds: • Pond #9 = 156 g/l (15.6%) lowest salinity pond visited • Ponds #11 & #12 = >280 g/l (>28%) • Crystallizer Channels 1 & 2 • Collected 500 mls water samples for inoculation/isolation and molecular purposes; stored at 4oC

  8. Estimate of Abundance (MPN)

  9. Isolation of Hosts

  10. Isolation of Hosts

  11. Viral Isolation Crystallizer Channel 2-B (C2- B) Top agar overlay method: 1ml isolated cells + 1ml pre-centrifuged H20 sample + 4mls top agar

  12. Summary of All Viral Isolates to Date • Crystallizer Channel 2 – high concentration • Pond 11 – one plaque • Pond 12 – one plaque

  13. Next Steps • Molecular/phylogenetic analysis of initial water samples and of isolates; • Screen for more lytic phages and induce for temperate viruses (antibiotic treatment, UV irradiation, salinity shock); • Purify and grow up current viral isolates • Phage-host physiological experiments • Phage characterization • Visit ESSA this fall (??) and collect water for future PFGE viral community analysis.

  14. Early Microbial Ecosystems Research Group Acknowledgments (aka, the logo page) • Jesse Dillon, advisor, Cal State Long Beach Paul Ngo, undergraduate assistant Lamine Diallo, undergraduate assistant • Funding: NSF Minority Postdoctoral Fellowship • NASA EMERG • Government of Mexico & ESSA

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