Gladstone Investment (GAIN) 10-Q 2020 Q1 Quarterly 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 Acting Principal 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 Quarterly Report onForm 10-Q for the quarter ended June 30, 2019(“Form 10-Q”), 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-Q fairly presents, in all material respects, the financial condition and results of operations of the Company.

Date: July 31, 2019

/s/ Nicole Schaltenbrand

Nicole Schaltenbrand

Acting Principal Financial Officer