The protesters gathered on Friday near the island where the production is being filmed and raised banners saying: "Stop the Big Brother programme" and "We disapprove of Big Brother".
The crowd included MPs Muhammad Khalid, Adil Maawda and Issa al-Mutawa, while some women also took part in the peaceful two-hour demonstration.
"We called for this demonstration to protest the programme, and this expresses the feelings of Bahrainis," Abd Allah Hashim, the head of Al-Muharraq Civil Society, was quoted as saying in Friday's Akhbar Al-Khaleej newspaper.
The show has angered Islamist MPs in the tiny Gulf state, seven of whom have signed a demand to question Information Minister Nabil Al-Hamr in parliament over the programme.
The reality programme has also
been controversial in the West
"We do not agree that the programme be filmed in Bahrain because it goes against our traditions. We support any development and tourism project provided it is not at the expense of our values and our traditions," Khalid told AFP on Wednesday.
The Middle East Broadcasting Centre (MBC) television channel is producing Big Brother at a villa in a resort called Amwaj on Muharraq island, the second largest in the Bahrain archipelago, and airing it nightly on its MBC2.
Twelve young men and women from across the Arab world are living in the villa in separate quarters, but meet in the lounge, kitchen and garden.
Film of their daily life and interaction imitates, in a more restricted way, scenes which angered traditionalists in the West when the programme first appeared in the Netherlands in 1999 and later in Britain, the US and France.
An information ministry official has called the demand to question the minister "unjustified", noting the production was not domestic under Bahrain state television, but made by MBC, a Saudi-owned company headquartered in Dubai.