White blood cell profile data for Chapter 3 of PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis

White blood cell profile data from Chapter 3 of the PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis.

Temperature variability, and in particular temperature decreases, can increase susceptibility of amphibians to infections by the fungus Batrachochytrium dendrobatidis (Bd). However, the effects of temperature shifts on the immune systems of Bd-infected amphibians are unresolved. We acclimated frogs to 16°C and 26°C (baseline), simultaneously transferred them to an intermediate temperature (21°C) and inoculated them with Bd (treatment), and tracked their infection levels and white blood cell profiles over six weeks. Study animals were captive-bred, juvenile Litoria caerulea.

For a separate initial study, forty-six individuals were raised in temperature-controlled rooms for 24 months and maintained on a natural photoperiod in individual enclosures. Twenty-six individuals were housed in a 16°C room; twenty individuals were housed in a 26°C room. For the study, we removed frogs from the temperature-controlled rooms, immediately divided them into temperature-controlled chambers set to 21°C, and administered Bd inoculations in the chambers. To ensure infection, we inoculated frogs on two consecutive days. On each day, we prepared a zoospore suspension of 6 x 105 zoospores per ml. To inoculate, we placed each frog into an individual 250-ml plastic container and added 5 ml (enough to cover the bottom of the container) of zoospore inoculant or sham inoculant to each container. We left frogs in inoculant baths for 12 hours per day. After each inoculation period, we returned frogs with their inoculant to individual permanent enclosures comprising 70 x 120 x 170 mm plastic containers lined with tap water-saturated paper towels.

To monitor Bd infection status and intensity over the course of the experiment, we swabbed frogs one week before removal from the temperature-controlled rooms (all tested Bd-negative) and 6 d, 14 d, 21 d, 29 d, 36 d, and 41 d post-inoculation. We determined the number of Bd zoospore genome equivalents per swab with a real-time quantitative PCR protocol.

Blood samples (0.1 ml) were collected by cardiac puncture with 26-ga needles one week before removing frogs from temperature-controlled rooms (baseline) and on days 6, 21, and 33. Blood smears were air-dried, stained with Wright’s stain, and observed by light microscopy at 100x. To perform a differential WBC count, we located the section of the blood smear with an even monolayer of cells (red blood cells as close together as possible without overlapping). We then counted and identified 200 WBCs per slide with a modified battlement technique. We identified WBCs as lymphocytes, neutrophils, monocytes, basophils, and eosinophils. We calculated WBC profiles for each frog by dividing the number of each cell type by the total number of cells counted.

This dataset shows our white blood cell counts. Column headings are explained below.

Frog = individual frog identifier

Start_temp = temperature of temperature-controlled room in which frog was maintained prior to study; cold = 16°C; hot = 26°C

Sul = frog snout-urostyle length

Treat_temp = Temperature of temperature-controlled chamber in which frog was maintained during study; medium = 21°C (treatment); cold = 16°C (sham-inoculated control); hot = 26°C (sham-inoculated control)

Day = day of experiment

Lymph = number of lymphocytes counted

Neut = number of neutrophils counted

Mon = number of monocytes counted

Eosin = number of eosinophils countedBas= number of basophils counted

Total_cells = total number of cells counted

Fields = number of microscope fields examined to reach 200 cells counted

Perc_lymph = proportion of counted cells that were lymphocytes

Perc_neut = proportion of counted cells that were neutrophils

Perc_mon = proportion of counted cells that were monocytesPerc_unk_bas = proportion of counted cells that were basophils

Perc_eosin = proportion of counted cells that were eosinophils

    Data Record Details
    Data record related to this publication White blood cell profile data for Chapter 3 of PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis
    Data Publication title White blood cell profile data for Chapter 3 of PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis
  • Description

    White blood cell profile data from Chapter 3 of the PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis.

    Temperature variability, and in particular temperature decreases, can increase susceptibility of amphibians to infections by the fungus Batrachochytrium dendrobatidis (Bd). However, the effects of temperature shifts on the immune systems of Bd-infected amphibians are unresolved. We acclimated frogs to 16°C and 26°C (baseline), simultaneously transferred them to an intermediate temperature (21°C) and inoculated them with Bd (treatment), and tracked their infection levels and white blood cell profiles over six weeks. Study animals were captive-bred, juvenile Litoria caerulea.

    For a separate initial study, forty-six individuals were raised in temperature-controlled rooms for 24 months and maintained on a natural photoperiod in individual enclosures. Twenty-six individuals were housed in a 16°C room; twenty individuals were housed in a 26°C room. For the study, we removed frogs from the temperature-controlled rooms, immediately divided them into temperature-controlled chambers set to 21°C, and administered Bd inoculations in the chambers. To ensure infection, we inoculated frogs on two consecutive days. On each day, we prepared a zoospore suspension of 6 x 105 zoospores per ml. To inoculate, we placed each frog into an individual 250-ml plastic container and added 5 ml (enough to cover the bottom of the container) of zoospore inoculant or sham inoculant to each container. We left frogs in inoculant baths for 12 hours per day. After each inoculation period, we returned frogs with their inoculant to individual permanent enclosures comprising 70 x 120 x 170 mm plastic containers lined with tap water-saturated paper towels.

    To monitor Bd infection status and intensity over the course of the experiment, we swabbed frogs one week before removal from the temperature-controlled rooms (all tested Bd-negative) and 6 d, 14 d, 21 d, 29 d, 36 d, and 41 d post-inoculation. We determined the number of Bd zoospore genome equivalents per swab with a real-time quantitative PCR protocol.

    Blood samples (0.1 ml) were collected by cardiac puncture with 26-ga needles one week before removing frogs from temperature-controlled rooms (baseline) and on days 6, 21, and 33. Blood smears were air-dried, stained with Wright’s stain, and observed by light microscopy at 100x. To perform a differential WBC count, we located the section of the blood smear with an even monolayer of cells (red blood cells as close together as possible without overlapping). We then counted and identified 200 WBCs per slide with a modified battlement technique. We identified WBCs as lymphocytes, neutrophils, monocytes, basophils, and eosinophils. We calculated WBC profiles for each frog by dividing the number of each cell type by the total number of cells counted.

    This dataset shows our white blood cell counts. Column headings are explained below.

    Frog = individual frog identifier

    Start_temp = temperature of temperature-controlled room in which frog was maintained prior to study; cold = 16°C; hot = 26°C

    Sul = frog snout-urostyle length

    Treat_temp = Temperature of temperature-controlled chamber in which frog was maintained during study; medium = 21°C (treatment); cold = 16°C (sham-inoculated control); hot = 26°C (sham-inoculated control)

    Day = day of experiment

    Lymph = number of lymphocytes counted

    Neut = number of neutrophils counted

    Mon = number of monocytes counted

    Eosin = number of eosinophils countedBas= number of basophils counted

    Total_cells = total number of cells counted

    Fields = number of microscope fields examined to reach 200 cells counted

    Perc_lymph = proportion of counted cells that were lymphocytes

    Perc_neut = proportion of counted cells that were neutrophils

    Perc_mon = proportion of counted cells that were monocytesPerc_unk_bas = proportion of counted cells that were basophils

    Perc_eosin = proportion of counted cells that were eosinophils

  • Other Descriptors
    • Descriptor

      This dataset is available as a comma-separated values (.csv) file.

    • Descriptor type Note
  • Data type dataset
  • Keywords
    • chytridiomycosis
  • Funding source
  • Research grant(s)/Scheme name(s)
  • Research themes
    Tropical Ecosystems, Conservation and Climate Change
    FoR Codes (*)
    • 06 - Biological Sciences
    SEO Codes
    Specify spatial or temporal setting of the data
    Temporal (time) coverage
  • Start Date
  • End Date
  • Time Period
    Spatial (location) coverage
  • Locations
    Data Locations

    Type Location Notes
    Attachment WBC_profile_data.csv
    The Data Manager is: Sasha Eden Greenspan
    College or Centre
    Access conditions Open: free access under license
  • Alternative access conditions
  • Data record size 1 file: 13.6 KB
  • Related publications
      Name Greenspan, Sasha E., Bower, Deborah S., Webb, Rebecca J., Berger, Lee, Rudd, Donna, Schwarzkopf, Lin, and Alford, Ross A. (2017) White blood cell profiles in amphibians help to explain disease susceptibility following temperature shifts. Developmental and Comparative Immunology, 77. pp. 280-286.
    • URL https://doi.org/10.1016/j.dci.2017.08.018
    • Notes Chapter 3 of PhD thesis published in Developmental and Comparative Immunology
  • Related websites
      Name
    • URL
    • Notes
  • Related metadata (including standards, codebooks, vocabularies, thesauri, ontologies)
  • Related data
      Name
    • URL
    • Notes
  • Related services
      Name
    • URL
    • Notes
    Citation Greenspan, Sasha Eden (2017): White blood cell profile data for Chapter 3 of PhD thesis: Thermal thresholds in the amphibian disease chytridiomycosis. James Cook University. https://doi.org/10.4225/28/5a0d1ac51a005