I haven't done any test on my server, but I will quote another SO answer:
Also, when [.htaccess is] enabled the server will take a potential performance hit. The reason is because, every server request, if .htaccess support is enabled, when Apache goes to fetch the requested file for the client, it has to look for a .htaccess file in every single directory leading up to wherever the file is stored. stackoverflow.com/a/29114826/6813732
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I haven't done any test on my server, but I will quote another SO answer: