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Objective To undertake a study on prevalence of Deg Nala disease in eastern states of India and to reproduce the disease in buffaloes by the Fusarium spp., isolated from the affected region. Methods During this investigation, a survey was conducted covering four states of eastern region to identify the Deg Nala cases as well as to isolate and characterize the causative agent(s). An experimental study was carried out to reproduce the disease in healthy male buffaloes (2–3 years age) by randomly dividing them into five groups (four in each group). Each individual group was fed with rice straw artificially infested with either of the two representative isolates of Fusarium oxysporum (F. oxysporum) (F01, F02) or representative reference strains of Fusarium equiseti (F. equiseti) (ITCCF-2470) and Fusarium moniliforme (F. moniliforme) (ITCCF-4821) for 30 days, whereas the control group was fed with normal rice straw only. Results A total of 658 Deg Nala cases were recorded and 12 Fusarium isolates were identified from the mouldy rice straw collected from these affected areas. The characterization of the isolates revealed three species viz., F. oxysporum, F. equiseti and F. moniliforme , among which F. oxysporum was predominant. The disease was artificially reproduced in three buffaloes in F01 group and one in F02 group within 20–23 days by feeding F. oxysporum infested rice straw which resembled the clinical symptoms and gross lesions of natural Deg Nala cases. Conclusions The field investigation and laboratory studies, including experimental production of Deg Nala disease suggest the possible involvement of mycotoxins. However, further investigations needs to be done to understand nature of the toxic factors involved in production of the Deg Nala disease.