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The film was originally announced for a 2000 release, but Full Moon's financial hardships delayed the production for decades. The studio eventually rebounded, and it was slated to be shot in 2020, but worldwide COVID lockdowns resulted in another production delay.

Filming was also originally set to take place in Albania before being re-located to Serbia. Croatia had also been considered for a shooting location.

When the film was announced for a 2000 release, filming at the time was to commence in Romania, akin to the previous Subspecies films, including its spin-off, Vampire Journals.

The first entry in the Subspecies franchise to not be filmed in Romania.

In a 2023 interview with Pop Horror, Ted Nicolaou spoke about getting the two leads to return after so many years: "Anders Hove is a dear friend of mine. After Subspecies (1991) where we had so many conflicts and very difficult production circumstances, we made it for Bloodstone: Subspecies II (1993) and Bloodlust: Subspecies III (1994), where there would be no drinking on the set and only drinking when we took off the make-up. I would join him and we would drink, and we became very close friends. And Denice Duff, we all stayed in touch over the years, and so we were all hoping that someday we would get to do Subspecies V. It didn't take convincing for Anders as much as it took cajoling him because he had sort of retired from the theater by this point and was not sure if he was still into acting, or if he could even come back to this role that he played so many years ago. And he played it so effectively. He was very nervous about approaching the film. But the thing with Anders is, once you put him in costume and in front of the camera, all of that anxiety disappears and he becomes a character that just from the first day of shooting, I was in awe of him."