Oamaru police said they were following several lines of inquiry after a witness saw a little blue penguin (kororā) run over and killed at the Esplanade in Oamaru.
The penguin was killed when "anti-social" vehicle activity was reported in the area.
Doc Dunedin operations manager Gabe Davies said "the death of this penguin appears to be a shockingly senseless act".
"The death appears to be caused by a vehicle and investigations are ongoing regarding the circumstances surrounding the death and persons responsible," he said.
Doc was working with Oamaru police and urging the public to come forward with any information that would help with the investigation into the death on September 28 in the Esplanade.
Sergeant Blair Wilkinson said police received a report that there were up to seven vehicles and a motorcycle being driven "anti-socially" in the Esplanade area at 9.15pm that day.
A witness described seeing a penguin run over and killed, he said.
Doc confirmed the witness retrieved the injured penguin, which was initially alive, but died soon after.
Forest & Bird Waitaki branch chairwoman Chloe Searle said the organisation was "incredibly disappointed" to hear of the penguin’s death.
"Most people in this community know how special those birds are to our local identity.
"Myself and other members are really heartbroken and disappointed. So much community effort has gone into helping the birds so it’s really sad to hear this," Ms Searle said.
The Oamaru Harbour area is home to hundreds of little penguins along with other wildlife, and boasts the popular tourist attraction, the Oamaru Penguins colony, where the species is protected.
Sergeant Tony Woodbridge said police were appealing to anybody who was part of the reported group of vehicles "to do the right thing".
"Come forward to speak to us. We’re also calling on the public to provide any information that could assist our investigation.
"We’d like to speak to anybody who was on or around the Esplanade between 8.30pm and 9.30pm on that Sunday," he said.
Mr Davies said investigators were interested in video, CCTV or mobile phone footage which could help determine what happened and provide evidence for a potential prosecution under the Wildlife Act.
"Kororā are such precious birds and are the world’s smallest penguins. They live and nest so close to an urban environment and are a popular tourist attraction.
"They are threatened and in decline and should be protected by the public and not put further at risk," Mr Davies said.
The maximum penalty for killing a protected species such as the kororā is two years imprisonment, a fine of $100,000 or both, he said.
Mr Davies said with more wildlife nesting and living next to coastal highways, it was very important for drivers to be more aware and to watch out for penguins, seals, sea lions and other species on the road.
"We are heading into a busy summer season and it’s shocking that these incidents keep occurring.
"Police are still investigating the deaths of three sea lions (pakake) which were shot near the mouth of the Waitaki River in August. All three had wounds consistent with being shot by a firearm."
Mr Davies said while the "appalling incidents" were rare, Doc received too many reports of people not acting in a respectful way towards New Zealand’s native wildlife.
• Call 0800 DOC HOT or police 105, reference number P063957021.
