This is the first time fecal prostaglandin F2α metabolite (PGFM) analysis and altrenogest were used to determine and maintain a pregnancy in a zoo-housed black-footed cat (BFC; Felis nigripes). The established pair had not produced offspring during the year since their arrival at Birmingham Zoo. Fecal samples were collected daily and analyzed by enzyme immunoassay for progestagen, estrogen metabolite, and PGFM concentrations. After a 4-mo separation, the pair was reintroduced, and breeding was reinitiated. Two pregnancies were suggested by sustained, elevated progestagen concentrations (mean >3 × baseline) but were confirmed by elevated PGFM concentrations (mean 4–25 × baseline) beginning about 1 mo after presumed or observed breeding. The first pregnancy was lost after ∼51 days, and altrenogest was administered (0.088 mg/kg/day) in order to help sustain the second pregnancy, which went to term, but the kitten did not survive. Ultrasonographic and radiographic evaluations of pregnancy were utilized during the second pregnancy to provide information on litter size.