Imbalance between sympathetic and parasympathetic inputs to the heart is strongly linked to cardiac dysfunction in type 2 diabetes. We recently showed via in vivo recordings that left cardiac sympathetic nerve activity (SNA) was increased in Zucker type 2 Diabetic Fatty (ZDF) rats. Importantly, in diabetes disturbances in heart rate regulation are a common dysfunction, however right cardiac SNA and parasympathetic nerve activity (PSNA) are unknown. Therefore, we aimed to directly measure right cardiac SNA and vagal PSNA in type 2 diabetes. 20-week old male diabetic ZDF rats (DM, n=6-9) and their non-diabetic (ND, n=6-7) littermates were anaesthetised with sodium pentobarbital intraperitonially (80mg/kg), and maintained by variable femoral vein infusion (~0.6mg/kg/min). The right cardiac sympathetic nerve and right parasympathetic vagal nerve were placed uncut over bipolar platinum recording electrodes. SNA and PSNA were recorded under baseline conditions and following intravenous injection of β-agonist isoproterenol (ISO; 1ug/kg). Animals were euthanised by overdose, and differences between groups were assessed via t-test. Right integrated cSNA was increased in DM (ND 1.7 ± 0.4 vs DM 6.0 ± 2.1 µV/s, p<0.05), in agreement with our measures of left cSNA, but despite reduced basal HR in diabetes. However, basal vagal PSNA firing rate was significantly increased in DM (ND 2.3 ± 1.1 vs DM 15.7 ± 5.6 Hz, p<0.05). Integrated cSNA was increased following ISO in ND animals, alongside increased heart rate, an effect significantly attenuated in DM (ND 21.0 ± 5.9 vs DM 3.9 ± 2.9 % change, p<0.05). Thus both right cSNA and PSNA are increased in DM at basal conditions, with most likely the PSNA changes being dominant because basal HR is lower in diabetes. Interestingly, the reduced reponsiveness of both SNA and PSNA to β-adrenergic stimulation suggests that severe impairment of autonomic control of the heart is likely a key contributor to the vast burden of cardiac dysfunction in type 2 diabetes.