Gladstone Investment (GAIN) 10-K 2017 FY Annual report

Exhibit 32.2

CERTIFICATION PURSUANT TO 18 U.S.C. SECTION 1350,

AS ADOPTED PURSUANT TO

SECTION 906 OF THE SARBANES-OXLEY ACT OF 2002

The undersigned, the Chief Financial Officer of Gladstone Investment Corporation (the “Company”), hereby certifies on the date hereof, pursuant to 18 U.S.C. §1350(a), as adopted pursuant to Section 906 of The Sarbanes-Oxley Act of 2002, that the Annual Report onForm 10-K for the fiscal year ended March 31, 2017(“Form 10-K”), filed concurrently herewith by the Company, fully complies with the requirements of Section 13(a) or 15(d) of the Securities Exchange Act of 1934, and that the information contained in theForm 10-K fairly presents, in all material respects, the financial condition and results of operations of the Company.

Date: May 15, 2017

/s/ Julia Ryan

Julia Ryan

Chief Financial Officer and Treasurer