April Fool's - a day later
My personal favorite prank -- which I've still never figured out how to top -- was about a dozen years ago. As I mention on my About page, I used to manage a DEC VAX system. Early on April 1 of some year in the late 80's, I plopped in some code in the global logon script so that when folks first powered up their VT terminals (this was back in the days of dumb terminals on most desks), everything displayed upside down and backwards. After 5 or 10 seconds it would then revert to normal. This was an insurance company, so it was a bit of a risk getting too radical with a prank, but the altered state of the display was short-lived enough that most folks got a good laugh after an initial bit of panic.
These days all I typically come up with are the standard lame pranks like putting a tiny piece of paper wrapped around someone's RJ45 plug when they return from a vacation so they think their network connection's toast -- or removing the mouse ball (if they're not using an optical mouse!).
I'm probably just not drinking enough coffee.
Update: OK, before I get too many emails, here's a routine that will turn a Windows display upside down; it's disabled by simply pressing the <Esc> key.