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2020 ; 1
(5
): e218-e225
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SARS-CoV-2 in fruit bats, ferrets, pigs, and chickens: an experimental
transmission study
#MMPMID32838346
Schlottau K
; Rissmann M
; Graaf A
; Schön J
; Sehl J
; Wylezich C
; Höper D
; Mettenleiter TC
; Balkema-Buschmann A
; Harder T
; Grund C
; Hoffmann D
; Breithaupt A
; Beer M
Lancet Microbe
2020[Sep]; 1
(5
): e218-e225
PMID32838346
show ga
BACKGROUND: In December, 2019, a novel zoonotic severe acute respiratory
syndrome-related coronavirus emerged in China. The novel severe acute respiratory
syndrome coronavirus 2 (SARS-CoV-2) became pandemic within weeks and the number
of human infections and severe cases is increasing. We aimed to investigate the
susceptibilty of potential animal hosts and the risk of anthropozoonotic
spill-over infections. METHODS: We intranasally inoculated nine fruit bats
(Rousettus aegyptiacus), ferrets (Mustela putorius), pigs (Sus scrofa
domesticus), and 17 chickens (Gallus gallus domesticus) with 10(5) TCID(50) of a
SARS-CoV-2 isolate per animal. Direct contact animals (n=3) were included 24 h
after inoculation to test viral transmission. Animals were monitored for clinical
signs and for virus shedding by nucleic acid extraction from nasal washes and
rectal swabs (ferrets), oral swabs and pooled faeces samples (fruit bats), nasal
and rectal swabs (pigs), or oropharyngeal and cloacal swabs (chickens) on days 2,
4, 8, 12, 16, and 21 after infection by quantitative RT-PCR (RT-qPCR). On days 4,
8, and 12, two inoculated animals (or three in the case of chickens) of each
species were euthanised, and all remaining animals, including the contacts, were
euthanised at day 21. All animals were subjected to autopsy and various tissues
were collected for virus detection by RT-qPCR, histopathology
immunohistochemistry, and in situ hybridisation. Presence of SARS-CoV-2 reactive
antibodies was tested by indirect immunofluorescence assay and virus
neutralisation test in samples collected before inoculation and at autopsy.
FINDINGS: Pigs and chickens were not susceptible to SARS-CoV-2. All swabs, organ
samples, and contact animals were negative for viral RNA, and none of the pigs or
chickens seroconverted. Seven (78%) of nine fruit bats had a transient infection,
with virus detectable by RT-qPCR, immunohistochemistry, and in situ hybridisation
in the nasal cavity, associated with rhinitis. Viral RNA was also identified in
the trachea, lung, and lung-associated lymphatic tissue in two animals euthanised
at day 4. One of three contact bats became infected. More efficient virus
replication but no clinical signs were observed in ferrets, with transmission to
all three direct contact animals. Mild rhinitis was associated with viral antigen
detection in the respiratory and olfactory epithelium. Prominent viral RNA loads
of 0-10(4) viral genome copies per mL were detected in the upper respiratory
tract of fruit bats and ferrets, and both species developed SARS-CoV-2-reactive
antibodies reaching neutralising titres of up to 1/1024 after 21 days.
INTERPRETATION: Pigs and chickens could not be infected intranasally by
SARS-CoV-2, whereas fruit bats showed characteristics of a reservoir host. Virus
replication in ferrets resembled a subclinical human infection with efficient
spread. Ferrets might serve as a useful model for further studies-eg, testing
vaccines or antivirals. FUNDING: German Federal Ministry of Food and Agriculture.