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While in much of the Western world snakes are feared, in the small, rural, mountainous town of Cocullo, in the middle of central Italy, snakes are annually collected and celebrated in a sacro-profane ritual. Every 1st of May, Serpari (snake catchers) capture and showcase dozens of non-venomous snakes to celebrate the ritual of San Domenico. In order to detect potential zoonotic pathogens within this unique epidemiological context, parasites and microorganisms of snakes harvested for the "festa dei serpari" ritual was investigated. Snakes (n = 112) were examined and ectoparasites collected, as well as blood and feces sampled. Ectoparasites were identified morpho-molecularly, and coprological examination conducted through direct smear and flotation. Molecular screenings were performed to identify parasites and microorganisms in collected samples (i.e., Mesostigmata mites, Anaplasma/Ehrlichia spp., Rickettsia spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Babesia/Theileria spp., Cryptosporidium spp., Giardia spp., Leishmania spp. and helminths). Overall, 28.5% (32/112) of snakes were molecularly positive for at least one parasite and/or microorganism. Endosymbiont Wolbachia bacteria were identified from Macronyssidae mites and zoonotic vector-borne bacteria (e.g., Rickettsia, Leishmania), as well as orally transmitted pathogens (i.e., Cryptosporidium, Giardia, Proteus vulgaris, Pseudomonas), were detected from blood and feces. Thus, given the central role of the snakes in the tradition of Cocullo, surveys of their parasitic fauna and associated zoonotic pathogens may aid to generate conservation policies to benefit the human-snake interactions, whilst preserving the cultural patrimony of this event. Author summary: The "festa dei serpari" is a unique sacro-profane ritual held each 1st of May in the small town of Cocullo, Central Italy. In this ceremony, dozens of non-venomous free-ranging snakes are captured by serpari (snake catchers) and showcased to thousands of pilgrims and tourists. Therefore, we aimed to assess the parasites and microorganisms of snakes within this unique epidemiological context to identify potential zoonotic pathogens. Snakes were examined and ectoparasites, blood and feces were collected, and morpho-molecular studies were performed. Overall, 28.5% (32/112) of snakes were positive for at least one parasite and/or microorganism. We identified new records of Mesostigmata mites as well as Leishmania tarentolae for the first time in Italian snakes. Importantly, we detected zoonotic microorganisms such as Rickettsia sp. in Aesculapian snake, as well as orally transmitted pathogens (i.e., Cryptosporidium, Giardia, Pseudomonas sp., Proteus vulgaris) from blood and feces of four species of snakes. Thus, snakes handled in this tradition may play a role in the zoonotic transmission of pathogens, given the contact with humans during this unique event. [ABSTRACT FROM AUTHOR]