key: cord-0965297-mdjwksou
authors: Gortázar, Christian; Barroso-Arévalo, Sandra; Ferreras-Colino, Elisa; Isla, Julio; de la Fuente, Gabriela; Rivera, Belén; Domínguez, Lucas; de la Fuente, José; Sánchez-Vizcaíno, José M.
title: Natural SARS-CoV-2 infection in kept ferrets, Spain
date: 2021-01-14
journal: bioRxiv
DOI: 10.1101/2021.01.14.426652
sha: 5ad8caa75a9ebf243fa99bb7ab8641a52ce1d392
doc_id: 965297
cord_uid: mdjwksou

We found SARS-CoV-2 RNA in 6 of 71 ferrets (8.4%) and isolated the virus from one rectal swab. Natural SARS-CoV-2 infection does occur in kept ferrets, at least under circumstances of high viral circulation in the human population. However, small ferret collections are probably unable to maintain prolonged virus circulation.

Natural infection of animals with severe acute respiratory syndrome coronavirus 2 25 (SARS-CoV-2) has been reported in pet cats and dogs, zoo felids, and mustelids belonging to the 26 subfamily mustelinae [1] . Among mustelids, natural SARS-CoV-2 infections have been recorded 27 in farmed American mink (Neovison vison), and sporadically in a wild mink sampled close to an 28 infected farm in Utah 1 and in a kept pet ferret (Mustela putorius furo) from an infected 29 household in Slovenia 2 . Ferrets are common laboratory models and experimental infections have 30 evidenced their susceptibility and ability to transmit the virus to other ferrets. SARS-CoV-2 is 31 shed up to 8 days post-infection (dpi) in nasal washes, saliva, urine, and feces and is effectively 32 transmitted to naive ferrets by direct contact and via the air [2, 3]. Experimentally infected ferrets 33 display either no clinical signs or exhibit elevated body temperature and loss of appetite [2, 4] . 34

Ferrets are common pets 3, 4, 5 , and are also used as work animals for rabbit control. 35

However, it remains unknown if SARS-CoV-2 circulates among kept ferret populations and if 36 ferrets, like farmed mink, could contribute to virus maintenance. 37

We studied 71 ferrets belonging to seven owners and used as working animals for rabbit 39 hunting in Ciudad Real province, central Spain. Group sizes ranged from four to 21 (mean 10). 40

Twenty ferrets belonging to groups 1 and 2 were re-sampled 66 days after initial sampling. were used as negative controls. Cultured cells were maintained at 37 °C with 5% CO 2 , with a 66 daily observation of virus-induced cytopathic effect (CPE) and cellular death. After 6 days, cell 67 cultures were frozen, thawed, and subjected to three passages with inoculation of fresh Vero E6 cells with the lysates as described above. SARS-CoV-2 molecular detection was performed by 69 RT-qPCR on the supernatants from every passage to confirm the presence/absence of the virus in 70 the cell culture. 71

We found SARS-CoV-2 RNA in swab samples from 6 of 71 ferrets (8.4%) ( Table 2) This study received funding from ISCIII, Spanish Government, grant number: 89 COV20/01385 and EFC was supported by a grant from Universidad de Castilla-La Mancha, 90

Spain. Table 1 . Primer sequences and amplified fragment sizes in base pairs.

Assessing the risks of SARS-CoV-2 in Wildlife

Infection and Rapid 111 Transmission of SARS-CoV-2 in Ferrets

SARS-CoV-2 is transmitted via contact and via the air between ferrets

Susceptibility of ferrets, cats, 116 dogs, and other domesticated animals to SARS-coronavirus 2

Detection of 119 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR

Shotgun 122 proteomics of SARS-CoV-2 infected cells and its application to the optimization of 123 whole viral particle antigen production for vaccines

Detection of environmental SARS-CoV-2 RNA in a high prevalence 127 setting in Spain. Transboundary and Emerging Diseases

SARS-CoV-2 and the human-animal interface: outbreaks on mink farms

Address for correspondence: Christian Gortázar, SaBio research group at IREC Instituto de 133

Investigación en Recursos Cinegéticos (CSIC-UCLM)

The authors have no conflicts of interest to declare. 93