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    Experimental results on the growth of Aplanochytrium (a sea fan parasite) cells over a temperature gradient conducted at the Harvell lab at Cornell University
    
  
  
    
    

Experimental results on the growth of Aplanochytrium (a sea fan parasite) cells over a temperature gradient conducted at the Harvell lab at Cornell University

Website: https://www.bco-dmo.org/dataset/3719
Data Type: experimental
Version: 1
Version Date: 2012-09-12

Project
» Influence of Temperature and Acidification on the Dynamics of Coral Co-Infection and Resistance (Climate_CoralDisease)
ContributorsAffiliationRole
Harvell, DrewCornell University (Cornell)Principal Investigator
Mydlarz, LauraUniversity of Texas at Arlington (UT Arlington)Co-Principal Investigator
Burge, ColleenCornell University (Cornell)Contact
Rauch, ShannonWoods Hole Oceanographic Institution (WHOI BCO-DMO)BCO-DMO Data Manager

Abstract
Experimental results on the growth of Aplanochytrium (a sea fan parasite) cells over a temperature gradient. Two types of assays were used in two trials to quantify Labyrinthulomycota cultures: cell counts using a hemocytometer and total protein concentration.


Dataset Description

Two types of assays were used in two trials to quantify Labyrinthulomycota cultures: cell counts using a hemocytometer and total protein concentration.


Acquisition Description

Sampling and Analytical Methodology:
In the first trial, a Labyrinthulomycota culture held at 22 degrees C was divided into 18 sub-cultures that were incubated at 15 degrees C, 20 degrees C, 25 degrees C, 30 degrees C, and 32 degrees C in triplicate for three days. Culture temperatures and incubation period were based on previous visual observations of Labyrinthulomycota growth, where over-growth of culture flasks occurs after three days at temperatures of 25 degrees C and higher. In the second trial, nine sub-cultures were incubated at 20 degrees C, 25 degrees C, and 30 degrees C in triplicate for three days.

In the second trial, total protein was also assessed. After three days, the media was poured off, rinsed once with 3 mLs of 0.22 um-filtered artificial sea water, and replaced with 3 mLs of 0.22 um-filtered artificial sea water. With a sterile wooden dowel, the bottom of each culture was scraped until no Labyrinthulomycota growth was visible on the bottom of the culture. 700 uL of each culture was placed in a bead beater and mixed at 300 rpm for 30 sec; 400 uL was set aside for protein assays and 300 uL for cell counts and held on ice until use.

Prior to counting using a hemocytometer, cells were vortexed for about 20 sec. In each culture, cells were counted in triplicate.

Total protein was extracted from each sample by adding 400 uL of extraction buffer (0.15 ug mL-1 DTT in Tris-HCl) to each tube. The contents of the tube were mixed and lysed for 2 minutes with the Fisherbrand disposable pestle grinder system, and incubated for 45 minutes on ice for extractions. Protein was measured using the DC protein kit (Bio-Rad), and read in triplicate using the Synergy HT multi-Detection microplate reader with KC4 software (Biotek Instruments, Vermont) at 750 nm.


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Related Publications

Burge, C., Douglas, N., Conti-Jerpe, I., Weil, E., Roberts, S., Friedman, C., & Harvell, C. (2012). Friend or foe: the association of Labyrinthulomycetes with the Caribbean sea fan Gorgonia ventalina. Diseases of Aquatic Organisms, 101(1), 1–12. doi:10.3354/dao02487
Results

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Parameters

ParameterDescriptionUnits
trialExperimental trial number (1 or 2; see Acquisition Description). unitless
tempIncubation temperature. degrees C
count_avgAverage of total cells per temperature. # cells (unitless)
count_avg_seStandard error of count_avg. # cells (unitless)
count_log10_avgAverage of log 10 transformed cell counts per temperature. # cells (unitless)
count_log10_avg_seStandard error of count_log10_avg. # cells (unitless)
protein_avgAverage protein concentration per temperature. mg protein per culture
protein_avg_seStandard error of protein_avg. mg protein per culture
protein_log10_avgAverage of log 10 transformed protein concentration per temperature. mg protein per culture
protein_log10_avg_seStandard error of protein_log10_avg. mg protein per culture
repReplicate. unitless
countTotal number of cells per culture. # cells (unitless)
proteinProtein concentration measured in milligrams of protein per culture. mg protein per culture
count_log10Total cells per culture, log 10 transformed. # cells (unitless)
protein_log10Protein concentration, log 10 transformed. mg protein per culture

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Deployments

lab_Harvell

Website
Platform
Cornell
Description
Harvell Lab at Cornell University. See more information on the lab's website.


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Project Information

Influence of Temperature and Acidification on the Dynamics of Coral Co-Infection and Resistance (Climate_CoralDisease)

Coverage: Florida Keys & Puerto Rico


This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).

Coral reef ecosystems are highly endangered by recent increases in temperature and by projected increases in ocean acidification. Although temperature has been identified as a driver of some coral disease outbreaks, nothing is known about direct effects of acidification on host immunity and pathogen virulence, or the potential for synergism with temperature. Natural coral populations often suffer from simultaneous infection by multiple pathogens that can also influence host immune responses, but co-infection dynamics have not been investigated in invertebrate systems lacking classical adaptive immunity. Changing climate will very likely influence the outcome of single and co-infection.

This project will investigate the influence of environmental stress on co-infection dynamics of the sea fan coral, Gorgonia ventalina, with a fungal pathogen, Aspergillus sydowii and a protist parasite, SPX. The goal is to identify the mechanisms through which multiple infections, temperature and acidification modify host resistance, leading to changes in within- and among-colony rates of disease spread.

The objectives of this project are to:
(1) Identify incidence and co-infection frequency of Aspergillus sydowii and SPX. Detailed field surveys of the two diseases will test the hypothesis that co-infection is significant, provide valuable information about drivers of aspergillosis, and will help to characterize an emerging new sea fan disease.
(2) Investigate how co-infection influences sea fan susceptibility, resistance, and within host disease dynamics. Through manipulative lab inoculation experiments we will test the hypothesis that single infections increase susceptibility to a second pathogen.
(3) Examine the effects of temperature increase and ocean acidification on pathogen virulence, on underlying host resistance, and on the dynamics of single and co-infections.

The hypotheses that acidification will increase pathogen virulence and host susceptibility will be tested in a temperature and pH controlled experimental system. This system will also allow the potential synergistic effects of temperature and acidification on host immunity and co-infection dynamics to be explored. The primary intellectual merit of the proposed work will be a greater understanding of how changing climate mediates co-infection and immunity in a non-model invertebrate. While fungal pathogens are primarily opportunistic, labyrinthulid protozoans are recognized as primary pathogens in shellfish. Even in shellfish, little is known about co-infections involving labyrinthulids, and these protists are entirely unstudied in corals.

Publications associated with this project:
Burge CA, Douglas N, Conti-Jerpe I, Weil E, Roberts S, Friedman CS & CD Harvell. (May 2012) Friend or foe: the association of Labyrinthulomycetes with the Caribbean sea fan, Gorgonia ventalina. Dis Aquat Org. 101:1-12. doi: 10.3354/dao02487

Burge CA, Mouchka, ME, Harvell, CD & S Roberts. (In review) Immune response of the Caribbean sea fan, Gorgonia ventalina exposed to an Aplanochytrium parasite as revealed by transcriptome sequencing.



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Funding

Funding SourceAward
NSF Division of Ocean Sciences (NSF OCE)

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This document is created by info v 4.1f 5 Oct 2018 from the content of the BCO-DMO metadata database.    2021-03-01  08:44:36