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. 2011 Jun 16;6(6):e21059. doi: 10.1371/journal.pone.0021059

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This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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To test the optimal sperm concentration for zebrafish in vitro fertilization, various working sperm concentrations were assessed. These test concentrations (cells/ml) included 10⁸ (N = 5), 10⁷(N = 21), 10⁶(N = 26), 10⁵(N = 25), 10⁴ (N = 25), and 10³ (N = 16) sperm to initiate fertilization in the dishes. The means were significantly different (P<0.05) between the groups that have different letters (ANOVA). This determined that 10⁶ cells/ml was the minimal in vitro sperm concentration needed to achieve maximum fertilization success, below this concentration the fertilization success declined.