The Pope's acceptance of mothers being able to breastfeed in church was endorsed a second time during a baptismal ceremony in the Sistine Chapel, where 33 infants were christened.
Sunday's service was the latest example of the progressive leadership style the current Pontiff has introduced into the Vatican since his appointment in 2013.
"You mothers give your children milk, and even now, if they cry because they are hungry, breastfeed them, don't worry," Pope Francis said, according to NBC News' translation of his remarks.
Last year, the Pope made it known that he has no issue with mothers breastfeeding during his ceremonies and even went on to say that they should not feel uncomfortable.
"If they are hungry, mothers, feed them, without thinking twice. Because they are the most important people here," he encouraged.
One mother reportedly took the Pope up on his offer, and attended to her restless baby.
While many people might ask why a mother wouldn't breastfeed in church, others may feel it’s immodest and distracting.
Previous research has revealed that only 29 per cent of Australians "strongly agree" that women should be encouraged to breastfeed in public.
In particular, breastfeeding in sacred sites such as churches remains more so of a sensitive issue.
The annual ceremony, marking the end of the Christmas season, is held on the first Sunday after the Epiphany on January 6 - an important date in the Catholic calendar, where children of Vatican employees are initiated into the Christian faith.