Madura cattle are the germplasm of native cattle on the verge of extinction because of crossbreeding. Therefore, the present study was aimed to determine the serum nerve growth factor (NGF) concentration as a predictor of fresh ejaculate fertility parameters in Madura bull candidates. Eleven Madura bull candidates used for frozen semen production were selected for the study. Blood samples were collected using a vacutainer from the jugular vein for analyzing serum NGF and testosterone levels. Meanwhile, semen collection was conducted using an artificial vagina for sperm motility, viability, and concentration assessment. Data were analyzed to determine the correlation among variables and the linear regression of NGF concentration to other significant variables. The result showed that NGF had a significant correlation (p<0.05) with testosterone levels, sperm motility, viability, and concentration. A significant correlation was observed between testosterone levels, sperm concentration, and sperm viability. The regression equation among significantly correlated variables was determined. For artificial insemination, suitable bull ejaculates for obtaining frozen semen should reach at least 2.12 ng/mL of NGF levels, with sperm viability, sperm concentration, and testosterone levels of more than 78.63%, 1,462.177 million/mL ejaculate, and 25.67 ng/mL, respectively. This is the first study to identify NGF as a predictor of male fertility in bull candidates of Madura cattle. Therefore, NGF levels could be used as a marker of male fertility in Madura bull cattle candidates. Thus, based on the minimum NGF levels, the ejaculate of Madura bull candidate that meets the requirements for frozen semen production could predict fertility.