This study aimed to investigation of the rumen juice transfounation either alone, or in combination with Bacillus subtilis natto, or premix upon the general health condition, productivity as well as upon some ruminal and biochemical parameters in after weaning Holstein heifers. The obtained results revealed significant increase in the body weight in the experimental group receiving ruminal juice plus premix, followed by the group received ruminal juice plus Bacillus subtilis natto, and ruminal juice alone compared with the control group. The immune responses, in terms of high leukocyte counts, lymphocytes, and granulocytes, were also significantly upregulated in the group that received ruminal juice plus Bacillus subtilis natto, followed by the group that received ruminal juice plus premix, and ruminal juice, respectively. Interestingly, groups treated with ruminal juice plus Bacillus subtilis natto, and ruminal juice plus premix had a significant improvement in the hemoglobin level, and the antioxidant capacity. Likely, such treatment groups had significant improvements in both total protozoal count, and eligibility for insemination. In conclusion, the obtained results of the present study strongly recommend the use of ruminal juice in a combination with Bacillus subtilis natto, or premix to improve the fertility indices, and immune responses in Holstein heifers.