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This retrospective cohort study examined the effect of birth order on neonatal morbidity and mortality in very preterm twins.Using 2005 to 2012 data from the Canadian Neonatal Network, very preterm twins born between 240/7 and 326/7 weeks of gestation were included. Odds of morbidity and mortality of second-born cotwins compared with first-born cotwins were examined by matched-pair analysis. Outcomes were neonatal death, severe brain injury (intraventricular hemorrhage grade 3 or 4 or persistent periventricular echogenicity), bronchopulmonary dysplasia, severe retinopathy of prematurity (ROP) (> stage 2), necrotizing enterocolitis (≥ stage 2), and respiratory distress syndrome (RDS). Multivariable analysis was performed adjusting for confounders.There were 6,636 twins (3,318 pairs) included with a mean gestational age (GA) of 28.9 weeks. A higher rate of small for GA occurred in second-born twins (10 vs. 6%). Mortality was significantly lower for second-born twins (4.3 vs. 5.3%; adjusted odds ratio: 0.75; 95% confidence interval [CI]: 0.59-0.95). RDS (66 vs. 60%; adjusted odds ratio: 1.40; 95% CI: 1.29-1.52) and severe retinopathy (9 vs. 7%; adjusted odds ratio: 1.46; 95% CI: 1.07-2.01) were significantly higher in second-born twins.Thus, while second-born twins had reduced odds of mortality, they also had increased odds of RDS and ROP.