The PM denied that his acquaintance with the owner of a large fishing business in the west Jutland town of Thybøron had influenced political decision making.
"I would immediately like to emphasise that it is my clear perception that the information I gave at the relevant [parliamentary] hearing [in November, ed.] was correct and complete," Rasmussen said in his opening remarks during Wednesday's consultation with MPs.
"I have not denied any relation to 'quota kings'. I have, however, clearly said that I have no relations that have in any way influenced the way I think," he said.
In August, the National Audit Office (Rigsrevisionen) published a report of the Ministry of Environment and Food's management of fishing quotas dating back to the early 2000s, taking in several ministers and both Liberal and Social Democrat-led governments.
The 2002 fisheries reform implemented transferable fishing quotas in an initiative aimed at preventing overfishing.
But last year's report stated that the Ministry's inadequate management of quota ownership had resulted in several fishermen exceeding their catch quotas.
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Administration of transferable fishing quotas by the ministry failed to prevent a limit on the number of so-called "quota kings", the report said.
The administration was "highly criticisable," the Audit Office wrote.
"I have probably met many fishermen with many different interests over the years," Rasmussen said at Thursday's hearing.
"But none of these relations have in any way exerted control over my or the government's fisheries policies. I also believe I am not the only one here with an acquaintance with a fisherman," he added.
The PM was summoned to a new parliamentary hearing over the issue after newspaper Ekstra Bladet reported he had been gifted a stay in a holiday villa to the value of 10,000 kroner (1,350 euros) by Thybøron fishing magnate John-Anker Hametner Larsen.
Larsen is also reported by the newspaper to be amongst fishing industry bosses and companies to have donated 190,000 kroner to charity Løkkefonden, which was founded by Rasmussen and is now run by his wife Sólrun Løkke Rasmussen.
The PM denied that the donations and villa stay had any influence on his politics.
He also rejected the suggestion he should at November's hearing have mentioned the holiday gift or having met Larsen at a function held by the latter's fishing company.
The November hearing was not focused on Rasmussen's relationship with fishing bosses, but his decision to remove fishing from the purview of Environment and Food minister Esben Lunde Larsen and transfer it to Karen Ellemann, the PM said.