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Listeriosis, a fatal food and environment borne disease of public health and veterinary importance, caused by Listeria spp. has not been explored in Pakistan. One hundred and fifty samples of fresh fruits and vegetables collected over a period of 12 months from various localities of Karachi city were screened according to the standard protocols. Suspected isolates were subjected to conventional methods for identification which included morphological, cultural, biochemical and serological tests. Out of 30 samples each of papaya, water melon and cantaloupe, and 15 each of cucumber, tomato, radish and carrot, Listeria monocytogenes was isolated from two samples of papaya and tomato each and one sample of water melon and cucumber each. It is inferred that fruits and vegetables, if not handled hygienically and served fresh, may be responsible for the transmission of listeriosis.