Toru Hashimoto, the hugely popular mayor of Osaka and a rising star on the national political stage, described the order to about 34,000 city workers earlier this month as "necessary for labour affairs management".
"As a matter of course, it will never happen that someone is promoted without any problem after ignoring a legitimate order from his or her superior," Hashimoto told reporters on Tuesday.
Body art is deeply taboo in Japan. Public baths, fitness centres and many other venues throughout the country usually ban customers with tattoos.
Most Osaka employees have filled out the survey -- with about 110 admitting to sporting a tattoo, most of them waste disposal workers -- but more than 500 others have not, according to media reports.
"Citizens feel uneasy or intimidated if they see tattoos (on workers) in services and it undermines trust in the city," Hashimoto has said.
The mayor reportedly became infuriated earlier this year after learning that a worker at a children's home threatened kids by showing them his tattoos.
Hashimoto -- who says he admires cockroaches and allegedly offered on air to impregnate a TV personality -- earlier said he would "turn down" American pop diva Lady Gaga or actor Johnny Depp if the tattooed pair applied for a job. (AFP)