Biochemical methane potential (BMP) assay was conducted at 35 degrees C to evaluate the effects of inoculum to substrate ratio (ISR) and substrate mix ratio between para-grass and pig manure co-digesting using different inocula. Rubber latex digester (RLD) inoculum showed higher methanogenic activity (41.4 mL CH4/gVS) than pig farm digester (PFD) inoculum (37.3 mL CH4/gVS). However, the maximum methane yields, occurred at the highest para-grass mix ratio (G), were 369.6, 437.6, 465.9 and 442.6 mL CH4/gTS(added) for RLD inoculum, versus 332.4, 475.0, 519.5 and 521.9 mL/gTS(added) for PFD inoculum at ISR 1, 2, 3, and 4, respectively. HPr, HBu and HVa appeared at higher G, corresponding to substrate's higher biodegradability. Response surface indicated that higher ISR and G had a significantly positive impact on methane yield. It suggested the use of higher ISR, i.e. 3 or 4, for BMP assay of these co-substrates. Dominant species of fermentative bacteria in each inoculum was tested by DGGE. (c) 2013 Elsevier Ltd. All rights reserved.