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Two types of modified nano-montmorillonite (MNM) were developed by ion-exchange reactions using two different surfactants; sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CETAB), to prepare MNMSDS and MNMCETAB, respectively. Both MNM types were on the nano-scale and had higher cation-exchange capacity values than NM clay. The MNMCETAB had the highest zeta potential (−27 mV) compared with the other clays. Effects of MNM types on in vitro ruminal batch culture fermentation, nutrient degradability, and methane (CH4) emission compared with monensin were evaluated in vitro using a semi-automatic gas production system. The experimental treatments were the control (0 supplementations), monensin (40 mg/kg DM), and NM (5 g NM/kg DM), and two levels of MNMSDS and MNMCETAB were supplemented at 0.05 (low) and 0.5 (high) g/kg DM to the control basal feed substrate. Among the experimental treatments, the high dose of both MNM types reduced (p < 0.01) CH4 production and ammonia concentrations compared with the control, while only MNMCETAB treatment tended to increase (p = 0.08) the truly degraded organic matter compared with monensin. All MNM treatments increased (p < 0.01) acetate molar proportions compared with monensin. The high MNMCETAB increased (p < 0.01) the in vitro ruminal batch culture pH compared with the control and monensin. The MNMCETAB supplemented at 0.5 g/kg DM is the most efficient additive to reduce CH4 emission with the advantage of enhancing the in vitro nutrient degradability of the experimental feed substrate. These results indicated that MNM could modulate the in vitro ruminal fermentation pattern in a dose- and type-dependent manner.