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Investigations have been launched into two trusts that work with animal parks in Kent.
The Charity Commission has opened formal inquiries into the Aspinall Foundation and Howletts Wild Animal Trust over "serious concerns about governance and financial management."
Howletts Wild Animal Trust runs Port Lympne in Hythe and Howletts in Canterbury. The Aspinall Foundation runs the conservation programmes.
They are being investigated separately.
The regulator began looking into Howletts in December 2019, due to concerns about the charity’s management of conflicts of interest and a related-party transaction.
A commission spokesman said: "We have today announced two new statutory inquiries, into The Aspinall Foundation and Howletts Wild Animal Trust.
"The respective charities have trustees in common, but are separate entities, and are subject to separate investigations.
"The trustees have co-operated fully with us since we first engaged with them last year.
"The Commission still has concerns about related party transactions, meaning that it is now necessary to examine these issues as part of a formal statutory inquiry."
The inquiry into Howletts will focus on concerns over the administration and management of the charity by trustees, and whether or not the trustees have complied with and fulfilled their duties and responsibilities under charity law.
Similarly, the trust began examining The Aspinall Foundation in July of last year, and since engaging with trustees in November, has "identified further issues with the charity's governance".
It will be investigating the same concerns as Howletts, and further querying whether or not there has been any "unauthorised trustee benefit".
A commission spokesman said that the opening of an inquiry is not a finding of wrongdoing.
"We remain firmly committed to our ethical and legal duties as a charitable body"
A spokesman for The Aspinall Foundation said that the trust is "co-operating fully" to resolve the Commission's concerns.
She continued: "We remain firmly committed to our ethical and legal duties as a charitable body.
"Our Trustees will continue to work openly and transparently with the Charity Commission to ensure best practice governance and compliance.
"No further comment will be issued until the investigation is complete."