To evaluate the signs and symptoms of dry eye in healthy pregnant women and investigate the effect of pregnancy on meibomian gland loss using non-contact meibography. Healthy pregnant women and age-matched healthy non-pregnant women were included in the study. Subjective symptoms were assessed using the Ocular Surface Disease Index (OSDI). The first and average non-invasive break-up times (first-NIBUT and avg-NIBUT, respectively) were determined, and the Schirmer test was applied. Non-contact meibography was performed. The study included a total of 62 women, 30 pregnant (study group) and 32 non-pregnant (control group). There was no significant difference between the groups in terms of OSDI score and the Schirmer test (p > 0.05). The mean first-NIBUT and avg-NIBUT values of the study group (13.1 ± 5.3 and 13.7 ± 4.5s) were significantly lower than the control group (16.0 ± 2.4 and 16.4 ± 1.5s) (p: 0.015 and p: 0.040, respectively). The mean meibomian gland losses in the upper and lower eyelids were significantly higher in the study group (16.9 ± 8.2% and 11.6 ± 10.2%) compared to the control group (9.7 ± 6.2% and 5.6 ± 4.7%) (p < 0.001 and p: 0.011, respectively). It is considered that loss of the meibomian glands may occur during pregnancy, and this may predispose pregnant women to evaporative dry eye. Although dry eye does not cause significant ocular surface symptoms in pregnancy, NIBUT and non-contact meibography can be considered as effective diagnostic methods for the detection of dry eye in pregnant women.