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Chytridiomycosis is a contributor to amphibian population declines. Diseased amphibians show symptoms of lethargy and loss of righting reflexes, likely due to an ion imbalance across the skin. However, it is possible developing zoosporangia release toxins that affect neuromuscular activity. Using Xenopus laevis as a model, we hypothesized that locomotor performance would be affected by injection of Bd supernatant factors. X. laevis were injected and then filmed performing a swimming escape response with high-speed cameras at 4 h, 24 h, and 1-week post-injection. Average maximum swimming velocity and escape latency were digitized using high-speed video. Despite no difference in escape velocity, there was a significant difference in escape latency 24 h post injection at both concentrations tested, 106 and 107 cell equivalents, though only differences at 106 cell equivalents/ml supernatant persisted 1 week post injection. Changes in specific locomotor function suggest that there may be neurotoxins present, though the potential neurotoxins may exhibit neural circuit specificity across escape behavior. This study provides a method to test more purified extracts to determine whether Bd produces neurotoxic factors that could enter the blood stream and alter locomotion during a natural skin infection.
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41503701 ???displayArticle.pmcLink???PMC12893034 ???displayArticle.link???Biol Open ???displayArticle.grants???[+]
IOS 2147467 National Science Foundation, BII 2120084 National Science Foundation, Belmont University
Fig. 1. Boxplots of the effect of three treatments on frog swim velocity (m/s) for all frogs. Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of 106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant. There was no significant difference in average maximum velocity between 107 cell count treatment (0.966 m/s±0.339) (N=12), 106 cell count treatment (1.020 m/s±0.339) (N=15), and sterile APBS (1.050 m/s±0.316) (N=16).
Fig. 2.
Boxplots of the effect of three treatments on frog escape latency (s) for all frogs. Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of 106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant. There was a significant difference in escape latency between 107 cell count treatment (0.3878 s±0.374) (N=12), 106 cell count treatment (0.381 s±0.332) (N=15), and sterile APBS (0.199 s±0.234) (N=16).
Fig. 3. Boxplots of the effect of three treatments and three trial times on escape latency (s) for all frogs. Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of 106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant.
Fig. 4. Boxplots of the effect of three trial times on frog swim velocity (m/s) of all frogs in all treatment groups (N=43). Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of 106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant. Average maximum velocity 4-h post infection (0.792 m/s±0.304) was significantly different from 24-h post injection (1.070 m/s±0.332) and 1-week post injection (1.190 m/s±0.284).
Fig. 5. Boxplots of the effect of three trial times on frog escape latency (s) of all frogs. Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of 106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant. Escape latency 4-h post infection (0.225 s±0.357) and 24-h post injection (0.257 s±0.292) were significantly different from 1-week post injection (0.434 s±0.277).
Fig. 6. Boxplots of the effect of three treatments and three trial times on frog average maximum swimming velocity (m/s) for all frogs. Boxes represent the first quartile, median, and third quartile. Solid line denotes median, dashed line denotes mean. A total of n=43 frogs were analyzed: 16 received the control (0.1 ml of sterile APBS), 15 received 0.1 ml of106 cell count equivalent supernatant, and 12 received 0.1 ml of 107 cell count equivalent supernatant.
