Australian osteopaths engage in multidisciplinary care and referrals with other health professionals, including general practitioners (GPs), for musculoskeletal care. This secondary analysis compared characteristics of Australian osteopaths who refer to, and receive referrals from, GPs with osteopaths who do not refer. The analysis was undertaken to identify pertinent characteristics that could contribute to greater engagement between Australian osteopaths and GPs. Data were from the Australian osteopathy practice-based research network comprising responses from 992 osteopaths (48.1% response rate). Osteopaths completed a practice-based survey exploring their demographic, practice, and clinical management characteristics. Backward logistic regression identified significant characteristics associated with referrals. Osteopaths who reported sending referrals (n = 878, 88.5%) to GPs were more likely than their non-referring colleagues to receive referrals from GPs (aOR = 4.80, 95% CI [2.62-8.82]), send referrals to a podiatrist (aOR = 3.09, 95% CI [1.80-5.28]) and/or treat patients experiencing degenerative spinal complaints (aOR = 1.71, 95% CI [1.01-2.91]). Osteopaths reporting receiving referrals (n = 886, 89.3%) from GPs were more likely than their non-referring colleagues to send referrals to GPs (aOR = 4.62, 95% CI [2.48-8.63]) and use the Medicare EasyClaim system (aOR = 4.66, 95% CI [2.34-9.27]). Most Australian osteopaths who report engaging in referrals with GPs for patient care also refer to other health professionals. Referrals from GPs are likely through the Chronic Disease Management scheme. The clinical conditions resulting in referrals are unknown. Further research could explore the GP-osteopath referral network to strengthen collaborative musculoskeletal care. The outcomes of this study have the potential to inform Australian osteopaths participating in advocacy, public policy and engagement with Australian GPs.