The message was part of a wider education programme about sexually-transmitted diseases and was aimed at reducing prejudice against sex workers.
Health Minister Alexandre Padilha said he had never endorsed the advert: "I do not think this is a message the ministry should be sending."
Critics in Brazil say the campaign glorified prostitution.
Mr Padilha said the message was being tested on the ministry's website, but it was not meant to be published.
"For as long as I remain in office, an ad like that will not be part of our campaigns," he told Agencia Estado.
The Brazilian government is highly praised for its Aids programme, which includes the distribution of free condoms and a pragmatic approach to the problem in an overwhelmingly Roman Catholic country.
But this is not the first time the Health Ministry's campaigns have attracted criticism.
Last year, the ministry dropped a video produced for the festive carnival period featuring a gay couple.
The current campaign of awareness to sexually-transmitted diseases will go ahead, without the "happy prostitute" reference.
It's entitled "Not ashamed of using a condom".
Brazil has experienced a boom in sex tourism in recent years.
With widespread poverty in the country, many boys and girls enter prostitution at a very young age.
The authorities at federal and state level have vowed to address the problem.
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