The video was posted to Twitter early Sunday morning.
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Later Sunday afternoon, the NYPD announced the officer who is under investigation for using a department speaker for political purposes has been suspended without pay.
NYPD Chief of Department Terence Monahan said that behavior will not be tolerated.
"When you wear our uniform it is imperative to remain apolitical," Monahan said on Twitter. "Behavior like this will not be tolerated and will be dealt with."
Earlier this week, Monahan and other NYPD officials held a press briefing to share their plans for Election Day in NYC.
At that news conference, Monahan vowed that NYPD officers would remain unbiased after an officer in Miami was spotted with a Trump face mask.
"Listen when we put on this uniform we are apolitical," Monahan said Tuesday. "We have no stance one way or the other, one role, and that's to keep people safe."
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NYPD Police Commissioner Dermot Shea said the video under investigation is unacceptable.
"One hundred percent unacceptable. Period," Shea said on Twitter. "Law Enforcement must remain apolitical, it is essential in our role to serve ALL New Yorkers regardless of any political beliefs. It is essential for New Yorkers to trust their Police."
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NYC Mayor Bill de Blasio said it will not be tolerated:
"Let me be clear: ANY NYPD Officer pushing ANY political agenda while on duty will face consequences. We will act fast here, and this will not be tolerated."
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The department, in its Patrol Guide, prohibits officers from "endorsing political candidates or publicly expressing personal views and opinions" about candidates while on duty or in uniform.
The NYPD put out a statement to say the matter is under investigation and the department plans to "come to an expedient resolution."
The Police Benevolent Association, the city's largest police union, threw its support behind Trump in August.
Trump, a Republican, has campaigned for reelection on a law-and-order agenda, portraying himself as a counterweight to the police reform protests that erupted across the U.S. in recent months.
Many police officers also support Trump, viewing him as more of an ally to their pro-police "Blue Lives Matter" and "Back the Blue" movements than his Democratic rival, former Vice President Joe Biden.
(The Associated Press contributed to this report.)