SARS-CoV-2 outbreak in lions, tigers, and hyenas at Denver Zoo.

Emily N Gallichotte1 Laura Bashor1 Katelyn Erbeck2 Lara Croft3 Katelyn Stache4 Jessica Long4 Sue VandeWoude1 James C Johnson4 Kristy L Pabilonia1,2 Gregory D Ebel1
Affiliations 4 institutions
  1. Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.
  2. Veterinary Diagnostic Laboratories, Colorado State University, Colorado, Fort Collins, USA.
  3. Morris Animal Foundation, Denver, USA.
  4. Denver Zoo Conservation Alliance, Denver, USA.

Abstract

In late 2019, SARS-CoV-2 spilled over from an animal host into humans, where it efficiently spread, resulting in the COVID-19 pandemic. Through both natural and experimental infections, we learned that many animal species are susceptible to SARS-CoV-2. Importantly, animals in close proximity to humans, including companion, farmed, and those at zoos and aquariums, became infected, and many studies demonstrated transmission to/from humans in these settings. In this study, we first review the literature of SARS-CoV-2 infections in tigers and lions and compare species, sex, age, virus and antibody detection assay, and types, frequency, and length of clinical signs, demonstrating broad heterogeneity among infections. We then describe a SARS-CoV-2 outbreak in lions, tigers, and hyenas at Denver Zoo in late 2021. Animals were tested for viral RNA (vRNA) for 4 months. Lions had significantly more vRNA in nasal swabs than both tigers and hyenas, and many individual lions experienced viral recrudescence after weeks of undetectable vRNA. Infectious virus was correlated with high levels of vRNA and was more likely to be detected earlier during infection. Four months post-infection, all tested animals generated robust neutralizing antibody titers. Animals were infected with Delta lineage AY.20 identical to a variant circulating at less than 1% in Colorado humans at that time, suggesting a single spillover event from an infected human spread within and between species housed at the zoo. Better understanding of epidemiology and susceptibility of SARS-CoV-2 infections in animals is critical to limit the current and future spread and protect animal and human health.IMPORTANCESurveillance and experimental testing have shown many animal species, including companion, wildlife, and conservatory, are susceptible to SARS-CoV-2. Early in the COVID-19 pandemic, big cats at zoological institutions were among the first documented cases of naturally infected animals; however, challenges in the ability to collect longitudinal samples in zoo animals have limited our understanding of SARS-CoV-2 kinetics and clearance in these settings. We measured SARS-CoV-2 infections over 4 months in lions, tigers, and hyenas at Denver Zoo and detected viral RNA, infectious virus, neutralizing antibodies, and recrudescence after initial clearance. We found lions had longer and higher levels of virus compared to the other species. All animals were infected by a rare viral lineage circulating in the human population, suggesting a single spillover followed by interspecies transmission. These data are important in better understanding natural SARS-CoV-2 spillover, spread, and infection kinetics within multiple species of zoo animals.

Supporting text Virus Host Location
COVID-19 467 cross-species transmission 75 spillover 105 Animals, Zoo 18 COVID-19 425 Disease Outbreaks 170 Lions 8 SARS-CoV-2 453 Tigers 8 Animals 1948 Antibodies, Viral 212 Colorado 4 Female 289 Humans 1440 Male 224 RNA, Viral 193

Evidence records

7 total
Zoonotic Surveillance
2 records · 2 evidence types
Evidence type
1 records
OVE8882
Key finding

SARS-CoV-2 viral RNA was detected in nasal swabs from lions, tigers, and hyenas at Denver Zoo, with lions showing higher viral RNA levels and episodes of recrudescence.

Virus
Host
Location
Supporting text

We measured SARS-CoV-2 infections over 4 months in lions, tigers, and hyenas at Denver Zoo and detected viral RNA, infectious virus, neutralizing antibodies, and recrudescence after initial clearance.

Method
viral RNA detection | vRNA assay
Sample type
nasal swabs
Geographic raw
Denver Zoo
Country inferred
USA
Evidence type
1 records
OVE8884
Key finding

Four months after SARS-CoV-2 infection, all tested lions, tigers, and hyenas at Denver Zoo developed robust neutralizing antibody titers, indicating serological response to exposure.

Virus
Host
Location
Not specified
Supporting text

Four months post-infection, all tested lions, tigers, and hyenas developed robust neutralizing antibody titers to SARS-CoV-2.

Method
neutralization assay | antibody titer measurement
Sample type
serum
Transmission Evidence
4 records · 2 evidence types
Evidence type
2 records
OVE8880
Key finding

A SARS-CoV-2 outbreak occurred in lions, tigers, and hyenas at Denver Zoo in late 2021, with viral RNA and antibody responses monitored for four months.

Virus
Host
Location
Not specified
Supporting text

We then describe a SARS-CoV-2 outbreak in lions, tigers, and hyenas at Denver Zoo in late 2021. Animals were tested for viral RNA (vRNA) for 4 months.

Method
viral RNA detection | neutralizing antibody titration | longitudinal sampling
Outbreak setting
Denver Zoo
Outbreak time
late 2021
Outbreak scale
zoo outbreak involving lions, tigers, and hyenas monitored for 4 months
OVE8881
Key finding

Zoo animals were infected with SARS-CoV-2 Delta lineage AY.20 identical to a rare variant in Colorado humans, suggesting a single human-to-animal spillover followed by spread among species.

Virus
Host
Location
Supporting text

Animals were infected with Delta lineage AY.20 identical to a variant circulating at less than 1% in Colorado humans at that time, suggesting a single spillover event from an infected human spread within and between species housed at the zoo.

Method
genomic sequencing | phylogenetic comparison | vRNA detection
Transmission direction
human-to-animal
Geographic raw
Colorado
Country inferred
USA
Outbreak setting
Denver Zoo
Outbreak time
late 2021
Outbreak scale
zoo outbreak involving multiple species
Evidence type
2 records
OVE8885
Key finding

A human-to-animal spillover of SARS-CoV-2 Delta lineage AY.20 occurred at the Denver Zoo, infecting lions, tigers, and hyenas after a human introduced the virus.

Virus
Host
Location
Supporting text

Animals were infected with Delta lineage AY.20 identical to a variant circulating at less than 1% in Colorado humans at that time, suggesting a single spillover event from an infected human spread within and between species housed at the zoo.

Method
viral genomic comparison | viral RNA detection | infectious virus isolation | antibody testing
Study design
natural outbreak investigation in zoo animals
Transmission direction
human-to-animal
Geographic raw
Colorado | Denver Zoo
OVE8886
Key finding

A SARS-CoV-2 outbreak in lions, tigers, and hyenas at Denver Zoo showed interspecies transmission among non-human hosts after a single human spillover.

Virus
Host
Location
Supporting text

We then describe a SARS-CoV-2 outbreak in lions, tigers, and hyenas at Denver Zoo in late 2021. All animals were infected by a rare viral lineage circulating in the human population, suggesting a single spillover followed by interspecies transmission.

Method
viral RNA detection | virus isolation | neutralizing antibody assay
Study design
outbreak investigation with longitudinal sampling
Transmission direction
animal-to-animal
Geographic raw
Denver Zoo
Country inferred
USA
Genomic Evolution
1 records · 1 evidence types
Evidence type
1 records
OVE8887
Key finding

Phylogenetic analysis showed that lions, tigers, and hyenas at the Denver Zoo were infected with the SARS-CoV-2 Delta lineage AY.20, genetically identical to a rare human variant in Colorado.

Virus
Host
Location
Not specified
Supporting text

Animals were infected with Delta lineage AY.20 identical to a variant circulating at less than 1% in Colorado humans at that time.

Analysis methods
phylogenetic characterization