The major disease that affects dairy industry in Pakistan is mastitis. Mastitis therapymostly fails due to pathological changes in udder, pharmacokinetics of drugs used and poor husbandrypractices. The present study is conducted to evaluate non-antibiotic therapy including Trisodium citrateand Vitamin C for the treatment of subclinical mastitis of cows and buffaloes. The product to be testedis composed of extemporaneous by mixing the Trisodium citrate 30gm and vitamin C 6gm. A fieldtrial on therapeutic efficacy of above product has been conducted on buffaloes and cows affected withsubclinical mastitis at Livestock Production Research Institute Bahadurnagar, Okara. Fifteen buffaloeswith 10 animals in treatment group (A1), 5 animals in control group (A2) fifteen cows with 10animals in treatment group (A3), 5 animals in control group (A4) are use in trial. Surf Field MastitisTest (SFMT) is used for grading of animals and treatment is given in both the groups of cattle andbuffalo. Buffalo group A1 and Cattle group A3 shows significantly higher (p<0.05) cure rates. Themilk pH in both the groups decreased up to 6.6. The efficacy of therapeutic protocol was calculated byAnalysis of Variance (ANOVA).