Two natural betaine sources; dehydrated condensed molasses fermentation solubles (Bet1) and Betafin®, a commercial anhydrous betaine extracted from sugar beet molasses and vinasses (Bet2); were used to investigate their impact on rumen fermentation parameters and lactation performance of lactating goats. Thirty-three lactating Damascus goats, with an average weight of 37 ± 0.7 kg and their age ranged from 22 to 30 months (2nd and 3rd lactation season), were divided into three groups, each group contained 11 animals. The control group (CON) was fed ration without betaine. While the other experimental groups were fed a control ration supplemented either with Bet1 or Bet2 to provide a 4 g betaine/kg diet. Results confirmed that betaine supplementation improved nutrient digestibility and nutritive value, and increased milk production and milk fat contents with both Bet1 and Bet2. Significant increases in concentration of ruminal acetate were observed in betaine-supplemented groups. Goats fed dietary betaine non-significantly recorded higher concentrations of short and medium-chain fatty acids (C4:0 to C12:0), and significant lower concentrations of C14:0 and C16:0 in milk. Also, both Bet1 and Bet2 non-significantly decreased the blood concentrations of cholesterol and triglycerides. Therefore, it could be concluded that betaine can improve the lactation performance of lactating goats and produce healthy milk with beneficial characteristics.