Objective: To examine the association between inter-twin delivery interval and short-term perinatal outcomes of the second twin after vaginal delivery of the first twin. Methods: Retrospective cohort study including twin pregnancies with a vaginal delivery of the first twin between January 2011 and September 2017 in a tertiary hospital in Melbourne, Australia. The main outcome measure was a composite of adverse neonatal outcome (at least one of perinatal death, admission to neonatal intensive care unit (NICU), endotracheal intubation, Apgar <7 at five minutes and cord lactate >4.0 mmol/L). Proportions of adverse outcomes for the second twin were compared between groups of intervals ≤ or >10 and ≤ or >30 min. Results: The composite adverse neonatal outcome occurred in 201 (58.2%) and a caesarean section occurred in seven cases (2%) of the 345 pregnancies included. Delivery interval was associated with higher cord lactate. Low Apgar scores were more frequent with intervals >30 min (17.9% vs 6.6%, P = 0.03), as well as caesarean section for the second twin (10.7% vs 1.3%, P = 0.01). Composite adverse outcome and admission to NICU were not significantly influenced by the delivery interval. Predictors of adverse outcome were gestational age, abnormal cardiotocography and breech delivery of the second twin. Conclusion: The inter-twin delivery interval is associated with higher rates of low Apgar scores and higher cord lactate for the second twin. These associations do not translate into higher rates of admission to NICU and their long-term clinical implications are uncertain.