Clinical symptoms and signs in hamsters during experimental infection with the SARS-CoV-2 virus (Coronaviridae: Betacoronavirus)

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Abstract

Introduction. At the beginning of December 2019, humanity has faced a new problem caused by coronavirus. In Hubei province of central China, epidemic events associated with severe primary viral pneumonia in humans began to develop. The isolated etiological agent was identified as a representative of Coronaviridae family. The global pandemic associated with the new coronavirus infection, acute respiratory syndrome type 2 (Severe acute respiratory syndrome 2, SARS-CoV-2), has become a challenge for humanity.

Objective. In our work, we assessed the replicative ability and pathogenesis of the SARS-CoV-2 virus in hamsters.

Materials and methods. Syrian hamsters (n=16) randomly divided into two groups were used in experiment. The first group was infected intranasally with the SARS-CoV-2 virus, strain SARS-CoV-2/human/KAZ/KZ_Almaty/2020 deposited in GenBank under number MZ379258.1. The second group remained as a control group. Clinical manifestations of the disease in hamsters were observed within 14 days. Samples were collected on days 3, 5, 7, 9, 12, and 14 postinfection. The obtained samples were tested for viral isolation in cell culture, histological examination and analysis of viral RNA by RT-PCR.

Results. SARS-CoV-2 virus isolates showed efficient replication in the lungs of hamsters, causing pathological lung lesions in animals infected intranasally. Clinical manifestations of the disease in hamsters infected with this virus were characterized by a decrease in temperature and body weight, wetness and ruffled fur, and frequent stroking of the nasal planum. High virus titers were observed following the virus isolation in cell cultures from nasal, oral swabs and lungs of animals infected intranasally. Pathological autopsy demonstrated pathological changes in the lungs. Moreover, transmission by airborne droplets has been established when a healthy hamster was kept together with animals infected using the intranasal method.

Conclusion. In conclusion, our study showed that the Syrian hamster model is a useful tool for studying the SARS-CoV-2 pathogenesis, as well as testing vaccine candidates against acute respiratory syndrome type 2.

About the authors

Moldir S. Tuyskanova

Research Institute for Biological Safety Problems; Al-Farabi Kazakh National University

Email: monica_94@list.ru
ORCID iD: 0000-0001-6565-082X

Master of Pedagogical Sciences, majoring in biology, junior researcher, Research Institute of Biological Safety Problems, Gvardeyskiy, Republic of Kazakhstan; Institute of the Ministry of Health of the Republic of Kazakhstan, Almaty, Republic of Kazakhstan

Kazakhstan, 080409, Gvardeyskiy; 050040, Almaty

Kuandyk D. Zhugunissov

Research Institute for Biological Safety Problems

Email: kuandyk_83@mail.ru
ORCID iD: 0000-0003-4238-5116

PhD, head of the laboratory Collection of microorganisms, Research Institute of Biological Safety Problems, Gvardeyskiy, Republic of Kazakhstan

Kazakhstan, 080409, Gvardeyskiy

Mehmet Ozaslan

Gaziantep University

Email: ozaslanmd@yahoo.com
ORCID iD: 0000-0001-9380-4902

professor, lecturer, Gaziantep University, Department of Biology

Turkey, 27310, Gaziantep

Balzhan S. Myrzakhmetova

Research Institute for Biological Safety Problems

Email: balzhan.msh@mail.ru
ORCID iD: 0000-0002-4141-7174

Candidate of Veterinary Sciences, Head of the Laboratory Especially Dangerous Infectious Diseases, Research Institute of Biological Safety Problems, Gvardeyskiy, Republic of Kazakhstan

Kazakhstan, 080409, Gvardeyskiy

Lespek B. Kutumbetov

Research Institute for Biological Safety Problems

Author for correspondence.
Email: lespek.k@gmail.com
ORCID iD: 0000-0001-8481-0673

professor, chief researcher, Research Institute of Biological Safety Problems, Gvardeyskiy, Republic of Kazakhstan

Kazakhstan, 080409, Gvardeyskiy

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2. Fig. 1. Indicators of thermometry and body weight measurements of infected animals. a – results of body temperature measurements in hamsters infected intranasally: max t °C, min t °C – maximum and minimum limits of normal body temperature for hamsters; «x» – on the third day, one hamster infected by the intranasal method died; ** – to establish infection with the virus during the joint keeping of animals, one hamster was transferred from the control group to the group of animals infected by the intranasal method instead of the dead hamster; b – results of the measurements of live body weight of hamsters.

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3. Fig. 2. Paraffin sections of hamster lungs. Pathohistological picture of the lung on the 3rd day after infection with SARS-CoV-2 virus. a – micrograph of the lung with diffuse damage to the alveoli, emphysema near the pleura and a small number of normally functioning alveoli; b – focal accumulation of microorganisms (indicated by an arrow) with contacting single neutrophils. Hematoxylin and eosin staining. Oс. ×10, lens ×4 (a); Oc. ×10, lens ×100 (under the immersion system) (b).

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4. Fig. 3. Paraffin sections of hamster lungs. Pathohistological picture of the lung on the 7th day after infection with SARS-CoV-2 virus. a – hyperemia, stasis and hemorrhagic necrosis in the vessels and bronchioles of the organ parenchyma; b – micropicture of restoration of the respiratory membrane, hyperemia and vasodilation of capillaries, infiltration by inflammatory cells. Hematoxylin and eosin staining. Oc. ×10, lens ×10 (a); Oc. ×10, lens ×40 (b).

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5. Fig. 4. Paraffin sections of hamster lungs. Pathohistological picture of the lung on the14th day after infection with SARS-CoV-2 virus. a – noticeable increase in the reducing zones of the lung parenchyma; b – bronchiole in a state of recovery with an internal content of red blood cells, microthrombi, hemorrhagic necrosis and diffuse infiltration of lymphoid tissue. Hematoxylin and eosin staining. Oc. ×10, lens ×4 (a); Oc. ×10, lens ×40 (b).

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6. Fig. 5. Paraffin sections of hamster lungs. Pathohistological picture of the lungs of hamsters from the control group. a – on the 7th day after the start of the experiment; b – on the 14th day after the start of the experiment; Hematoxylin and eosin staining. Oc. ×10, lens ×4 (a); Oc. ×10, lens ×40 (b).

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7. Fig. 6. Results of PCR testing of clinical and pathological materials from hamsters of the control group and those infected with the SARS-CоV-2 virus. a – nasal washes; b – oral washes; c – lung tissue.

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8. Fig. 7. Virus isolation from clinical and pathological materials from hamsters infected by various methods with the SARS-CoV-2 virus. a – nasal washes; b – oral washes; c – lung tissue.

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Copyright (c) 2023 Tuyskanova M.S., Zhugunissov K.D., Ozaslan M., Myrzakhmetova B.S., Kutumbetov L.B.

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