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Background  Although, use of animal species in disease treatment and culture practices is as ancient as that of plant species; however ethnomedicinal uses and cultural values of animal species have rarely been reported. Present study is the first report on the medicinal uses of mammals and bird species in Pakistan.    Methods  Questionnaires and semi-structured interviews were applied to collect qualitative and quantitative data from local informants ( N  = 109). Relative frequency of mention (RFM), fidelity level (FL), relative popularity level (RPL), similarity index (SI) and rank order priority (ROP) indices were used to analyzed the data.    Results  One hundred and eight species of animals, which include: 83% birds and 17% mammals were documented. In total 30 mammalian and 28 birds’ species were used to treat various diseases such as rheumatic disorders, skin infections and sexual weakness among several others. Fats, flesh, blood, milk and eggs were the most commonly utilized body parts.  Bos taurus, Bubalus bubalis, Capra aegagrus hircus, Felis domesticus, Lepus nigricollis dayanus  and  Ovis aries  (mammals) and  Anas platyrhynchos domesticus, Columba livia, Coturnix coturnix, Gallus gallus  and  Passer domesticus  (birds) were the highly utilized species. Medicinal and cultural uses of 30% mammals and 46% birds were reported for the first time, whereas 33% mammals and 79% birds depicted zero similarity with previous reports.    Conclusion  Present study exhibits significant ethnozoological knowledge of local inhabitants and their strong association with animal species, which could be helpful in sustainable use of biodiversity of the region. Additionally, in vitro and in vivo evaluation of biological activities in the mammalian and birds’ species with maximum fidelity level and frequency of mention could be important to discover animal based novel drugs.    Graphical Abstract  Some commonly used mammals and birds species of the study area

Ethnomedicinal and cultural practices of mammals and birds in the vicinity of river Chenab, Punjab-Pakistan