Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia

Bats are natural hosts of various coronaviruses (CoVs), including human CoVs, via an assumed direct zoonotic spillover or intermediate animal host. The present study aimed to investigate the circulation of CoVs in a bat colony in the Mediterranean region of Croatia. Guano and individual droppings fr...

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Main Authors: Dragan Brnić, Ivana Lojkić, Nina Krešić, Vida Zrnčić, Lea Ružanović, Tina Mikuletič, Martin Bosilj, Andrej Steyer, Tomislav Keros, Boris Habrun, Lorena Jemeršić
Format: Article
Language:English
Published: MDPI AG 2023-04-01
Series:Microorganisms
Subjects:
Online Access:https://www.mdpi.com/2076-2607/11/4/959
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author Dragan Brnić
Ivana Lojkić
Nina Krešić
Vida Zrnčić
Lea Ružanović
Tina Mikuletič
Martin Bosilj
Andrej Steyer
Tomislav Keros
Boris Habrun
Lorena Jemeršić
author_facet Dragan Brnić
Ivana Lojkić
Nina Krešić
Vida Zrnčić
Lea Ružanović
Tina Mikuletič
Martin Bosilj
Andrej Steyer
Tomislav Keros
Boris Habrun
Lorena Jemeršić
author_sort Dragan Brnić
collection DOAJ
description Bats are natural hosts of various coronaviruses (CoVs), including human CoVs, via an assumed direct zoonotic spillover or intermediate animal host. The present study aimed to investigate the circulation of CoVs in a bat colony in the Mediterranean region of Croatia. Guano and individual droppings from four bat species were sampled and tested with the E-gene sarbecovirus RT-qPCR, the pan-CoV semi-nested RT-PCR targeting the RdRp gene and NGS. Furthermore, bat blood samples were investigated for the presence of sarbecovirus-specific antibodies with the surrogate virus neutralization test (sVNT). The initial testing showed E-gene Sarebeco RT-qPCR reactivity in 26% of guano samples while the bat droppings tested negative. The application of RdRp semi-nested RT-PCR and NGS revealed the circulation of bat alpha- and betaCoVs. Phylogenetic analysis confirmed the clustering of betaCoV sequence with SARS-CoV–related bat sarbecoviruses and alpha-CoV sequences with representatives of the <i>Minunacovirus</i> subgenus. The results of sVNT show that 29% of bat sera originated from all four species that tested positive. Our results are the first evidence of the circulation of SARS-CoV–related coronaviruses in bats from Croatia.
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spelling doaj.art-021e72f849fa4e89b918e57f6f78afff2023-11-17T20:33:00ZengMDPI AGMicroorganisms2076-26072023-04-0111495910.3390/microorganisms11040959Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from CroatiaDragan Brnić0Ivana Lojkić1Nina Krešić2Vida Zrnčić3Lea Ružanović4Tina Mikuletič5Martin Bosilj6Andrej Steyer7Tomislav Keros8Boris Habrun9Lorena Jemeršić10Croatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaCroatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaCroatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaCroatian Biospeleological Society, Demetrova 1, 10000 Zagreb, CroatiaCroatian Biospeleological Society, Demetrova 1, 10000 Zagreb, CroatiaInstitute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, SloveniaNational Laboratory of Health, Environment and Food, Grablovičeva 44, 1000 Ljubljana, SloveniaNational Laboratory of Health, Environment and Food, Grablovičeva 44, 1000 Ljubljana, SloveniaCroatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaCroatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaCroatian Veterinary Institute, Savska cesta 143, 10000 Zagreb, CroatiaBats are natural hosts of various coronaviruses (CoVs), including human CoVs, via an assumed direct zoonotic spillover or intermediate animal host. The present study aimed to investigate the circulation of CoVs in a bat colony in the Mediterranean region of Croatia. Guano and individual droppings from four bat species were sampled and tested with the E-gene sarbecovirus RT-qPCR, the pan-CoV semi-nested RT-PCR targeting the RdRp gene and NGS. Furthermore, bat blood samples were investigated for the presence of sarbecovirus-specific antibodies with the surrogate virus neutralization test (sVNT). The initial testing showed E-gene Sarebeco RT-qPCR reactivity in 26% of guano samples while the bat droppings tested negative. The application of RdRp semi-nested RT-PCR and NGS revealed the circulation of bat alpha- and betaCoVs. Phylogenetic analysis confirmed the clustering of betaCoV sequence with SARS-CoV–related bat sarbecoviruses and alpha-CoV sequences with representatives of the <i>Minunacovirus</i> subgenus. The results of sVNT show that 29% of bat sera originated from all four species that tested positive. Our results are the first evidence of the circulation of SARS-CoV–related coronaviruses in bats from Croatia.https://www.mdpi.com/2076-2607/11/4/959SARS-CoVbatsarbecovirusRT-qPCRNGSsVNT
spellingShingle Dragan Brnić
Ivana Lojkić
Nina Krešić
Vida Zrnčić
Lea Ružanović
Tina Mikuletič
Martin Bosilj
Andrej Steyer
Tomislav Keros
Boris Habrun
Lorena Jemeršić
Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
Microorganisms
SARS-CoV
bat
sarbecovirus
RT-qPCR
NGS
sVNT
title Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
title_full Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
title_fullStr Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
title_full_unstemmed Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
title_short Circulation of SARS-CoV–Related Coronaviruses and Alphacoronaviruses in Bats from Croatia
title_sort circulation of sars cov related coronaviruses and alphacoronaviruses in bats from croatia
topic SARS-CoV
bat
sarbecovirus
RT-qPCR
NGS
sVNT
url https://www.mdpi.com/2076-2607/11/4/959
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