He returned to Florence to practice medicine, while also becoming personal physician to Grand Princess Violante di Baviera.
He had a wide range of interests and contributed poetry apart from participating in learned societies, founding the Societa Botanica Florentina along with other associates.
He wrote a pamphlet in 1725 suggesting that perennial springs were fed by sea waters through underground channels.
In 1731 he began to assemble a private natural history museum, the holding of which he catalogued in 1742, as Index Testarum Conchyliorum, quae adservantur in Museo Nicolai Gualtieri (translation: List of the shells of shellfish which are preserved in the museum of Niccolò Gualtieri) with copper plate engravings by Giuseppe Menabuoni and Antonio Pazzi.
His collections were acquired by Stefano Lorena and are now deposited at the Museo storia naturale di Pisa.