After being appointed as Roberto Mancini's replacement, Spalletti's hopes of getting off to a winning start against North Macedonia were dented by Enis Bardhi, who scored an 81st-minute free-kick to cancel out Ciro Immobile's opener.
In addition to the disappointing result, Spalletti received two injury setbacks in the contest after defender Gianluca Mancini and attacker Matteo Politano were forced off due to thigh and calf problems respectively.
The pair have subsequently withdrawn from the squad, and while Spalletti opted against replacing Mancini, Bologna winger Riccardo Orsolini has been called up as Politano's replacement.
Orsolini is not expected to start Tuesday's crucial encounter against second-placed Ukraine, but Italy will have to bring two players into the side in Mancini and Politano's absence.
Atalanta youngster Giorgio Scalvini is expected to be brought into the side to form a central defensive partnership with Alessandro Bastoni, and the pair will offer protection to Paris Saint-Germain goalkeeper Gianluigi Donnarumma.
Giovanni Di Lorenzo, who helped Napoli and Spalletti win the Serie A title last season, is set to retain his spot on the right side of defence, while Federico Dimarco will continue to operate in the left-back role.
Nicolo Barella will earn his 47th cap as he continues to close in on a half-century of international appearances, and the Inter Milan midfielder will be joined in the middle of the park by Roma's Bryan Cristante and Newcastle United's summer additions Sandro Tonali.
After scoring his 16th international goal, Italy captain Immobile will continue to lead the line alongside his Lazio teammate Mattia Zaccagni and Aston Villa's summer recruit Nicolo Zaniolo, who is expected to replace the injured Politano.
Italy possible starting lineup:
Donnarumma; Di Lorenzo, Scalvini, Bastoni, Dimarco; Barella, Cristante, Tonali; Zaniolo, Zaccagni, Immobile