Recovery of neurula cells after cryopreservation.

<p>Post-thaw recovery of cell viability (live/dead staining) expressed as mean number of viable cells per thousand counted (left hand column at each time) and the number of replicates with viable cells (right hand column at each time) for each treatment group. [CP] = cryoprotectant concentration.</p

Mean viability of cells from gastrulae and neurulae after 24 hours of incubation in various media/cryoprotectant combinations.

<p>(<b>A</b>) gastrulae; (<b>B</b>) neurulae. cm dmso = CM containing DMSO, sar dmso = SAR containing DMSO, cm glyc = CM containing glycerol, sar glyc = SAR containing glycerol. Each mean, n = 6 (2 embryos from each of 3 spawns); error bar = pooled standard error.</p

<i>Limnodynastes peronii</i> embryos and embryonic cells.

<p>(<b>A</b>) Late gastrula (Gosner Stage 12). (<b>B</b>) Neurula (Gosner Stage 15). (<b>C</b>) <i>L. peronii</i> embryos continuing normal development 24 hours after removal from holding at 8°C. (<b>D</b>) Post thaw viable and non-viable neurula cells determined with trypan blue live (L) - dead (D) staining; 400× magnification.</p

Mean viability of cells from gastrulae and neurulae incubated in various media/cryoprotectant combinations for 24 hours.

<p>(<b>A–D</b>) gastrulae; (<b>E–H</b>) neurulae. CM = culture medium; SAR = Simplified Amphibian Ringer; dmso = DMSO; glyc = glycerol; numbers 5–20 indicate % v/v concentration of cryoprotectant in media; control = 0% v/v cryoprotectant. Each mean, n = 6 (2 embryos from each of 3 spawns); error bar = pooled standard error.</p

Recovery of gastrula cells after cryopreservation.

<p>Post-thaw recovery of cell viability (live/dead staining) expressed as mean number of viable cells per thousand counted (left hand column at each time) and the number of replicates with viable cells (right hand column at each time) for each treatment group. [CP] = cryoprotectant concentration.</p

Effect of NaCl on developing zoosporangia.

<p>The mean (±SE) number of developing <i>Batrachochytrium dendrobatidis</i> zoosporangia counted within an ocular frame at 400× magnification when grown for 11 days in liquid media with varying salt concentrations. Different letters above bars indicate significant differences were found between groups.</p

Effect of NaCl on chytrid zoospores.

<p>The mean (±SE) number of <i>Batrachochytrium dendrobatidis</i> zoospores that were counted in or moving through an ocular frame at 400× magnification after 11 days growth in liquid media with varying salt concentrations. Different letters above bars indicate significant differences were found between groups.</p

Effect of NaCl on survival rates.

<p>Survival curves for juvenile <i>Litoria peronii</i> inhabiting tanks with waterbodies of varying salt concentrations (0–5 ppt) following inoculation with <i>Batrachochytrium dendrobatidis</i>. Death was assumed following the expression of lethargy and slow righting reflex typical of the terminal stages of chytridiomycosis. Different letters above lines indicate significant differences were found between groups.</p

Effect of NaCl on mature zoosporangia.

<p>The mean (±SE) number of mature <i>Batrachochytrium dendrobatidis</i> zoosporangia counted within an ocular frame at 400× magnification when grown for 11 days in liquid media with varying salt concentrations. Different letters above bars indicate significant differences were found between groups.</p

Effect of NaCl on infection load.

<p>The number of <i>Batrachochytrium dendrobatidis</i> genomic equivalents detected on epidermal swabs from juvenile <i>Litoria peronii</i> housed in tanks with water bodies at varying salt concentrations. Different letters indicate significant differences were found between groups.</p