We conducted a cross-sectional survey of 760 Australian layer chicken farms to identify risk factors associated with the closure of poultry farms. Address data and operating status were obtained for 734 farms and more detailed responses on farm size, structure and management practices from 317 farms (41.7%). The overall prevalence of closed farms was 36.4%. Increased risk of closure was associated with independent ownership [odds ratio (OR) = 2.22, P < 0.01], location within 50 km of the centre of a state capital city (OR = 1.66, P < 0.01), being a layer farm that reared its own pullets (OR = 2.18, P < 0.01) and location within Queensland or South Australia (OR = 3.10, 3.56; both P < 0.01). Analysis of the subset of farms for which more detailed information was available identified location in South Australia (OR = 9.23, P < 0.01), use of cages (OR = 4.36, P < 0.01) and sheds with only natural ventilation (OR = 11.00, P < 0.02) as risk factors associated with a greater risk of farm closure. Our survey found that closed farms were typically independently owned, cage-layer farms located in peri-urban regions near major cities and utilising sheds without assisted ventilation.