Abstract: Background and Objective: Avian colibacillosis is associated with heavy economic losses to the poultry industry and generally indicative of immunosuppression in the birds. It thus warrants rapid and accurate diagnostic approaches. Therefore, the present study was aimed to assess the pathological and pharmacochemical status of broiler chickens affected naturally with colibacillosis in Kashmir valley. Methodology: Commercial broiler chickens reared in Ganderbal District of Kashmir valley were used in this study. The samples included the diseased or dead birds received from various poultry farms. Gross pathological, histopathological, histoenzymatic and pharmacochemical observations were recorded. Results: On necropsy, among lymphoid organs, the major changes were observed in spleen and bursa. Histopathologically, lymphoid depletion was observed in bursa, thymus, spleen and caecal tonsils. No significant change in enzymatic activity in terms of acid phosphatase and alkaline phosphatase was observed in the lymphoid organs, with the exception of very mild enzyme activity in spleen. Pharmacochemical analysis showed that the caecal glandular epithelium, bursal plica epithelium and alveolar epithelium of harderian gland were positive for acid mucopolysaccharides with no signs of acid mucin in any other lymphoid organ. Conclusion: In conclusion, laboratory diagnosis is an important asset and proper lab diagnosis would serve as an important tool in preventing the economic losses incurred as a result of colibacillosis in poultry production.