The pathology of co-infection with Usutu virus and Plasmodium spp. in naturally infected Eurasian blackbirds (Turdus merula)

doi:10.1016/j.onehlt.2023.100534

. 2023 Apr 1:16:100534.

doi: 10.1016/j.onehlt.2023.100534. eCollection 2023 Jun.

The pathology of co-infection with Usutu virus and Plasmodium spp. in naturally infected Eurasian blackbirds (Turdus merula)

Gianfilippo Agliani¹, Giuseppe Giglia¹, Erwin de Bruin¹, Tjomme van Mastrigt^{2

3

4}, Rody Blom⁵, Reina S Sikkema^{3

6}, Marja Kik^{1

7}, Marion P G Koopmans⁶, Andrea Gröne^{1

7}, Judith M A Van den Brand^{1

7}

Affiliations

¹ Division of Pathology, Department of Biomolecular Health Sciences, Utrecht University, Utrecht, the Netherlands.
² Department of Animal Ecology, Netherlands Institute of Ecology (NIOO-KNAW), Wageningen, the Netherlands.
³ Vogeltrekstation - Dutch Centre for Avian Migration and Demography (NIOO-KNAW), Wageningen, the Netherlands.
⁴ Wildlife Ecology and Conservation Group, Wageningen University & Research, Wageningen, the Netherlands.
⁵ Laboratory of Entomology, Plant Sciences Group, Wageningen University & Research, Wageningen, the Netherlands.
⁶ Department of Viroscience, Erasmus Medical Center, Rotterdam, the Netherlands.
⁷ Dutch Wildlife Health Centre, Utrecht University, Utrecht, the Netherlands.

PMID: 37363251
PMCID: PMC10288036
DOI: 10.1016/j.onehlt.2023.100534

The pathology of co-infection with Usutu virus and Plasmodium spp. in naturally infected Eurasian blackbirds (Turdus merula)

Gianfilippo Agliani et al. One Health. 2023.

. 2023 Apr 1:16:100534.

doi: 10.1016/j.onehlt.2023.100534. eCollection 2023 Jun.

Authors

Affiliations

¹ Division of Pathology, Department of Biomolecular Health Sciences, Utrecht University, Utrecht, the Netherlands.
² Department of Animal Ecology, Netherlands Institute of Ecology (NIOO-KNAW), Wageningen, the Netherlands.
³ Vogeltrekstation - Dutch Centre for Avian Migration and Demography (NIOO-KNAW), Wageningen, the Netherlands.
⁴ Wildlife Ecology and Conservation Group, Wageningen University & Research, Wageningen, the Netherlands.
⁵ Laboratory of Entomology, Plant Sciences Group, Wageningen University & Research, Wageningen, the Netherlands.
⁶ Department of Viroscience, Erasmus Medical Center, Rotterdam, the Netherlands.
⁷ Dutch Wildlife Health Centre, Utrecht University, Utrecht, the Netherlands.

PMID: 37363251
PMCID: PMC10288036
DOI: 10.1016/j.onehlt.2023.100534

Abstract

Usutu virus (USUV) is a mosquito-borne zoonotic flavivirus causing mortality in Eurasian blackbirds (Turdus merula) in Europe. In dead blackbirds, avian malaria co-infection due to mosquito-borne hemosporidians (e.g., Plasmodium spp.) has been reported. In humans, a similar co-infection of a flavivirus, Dengue virus, and Plasmodium spp. is causing increased severity of clinical disease. Currently, the effects of co-infection of arboviruses and hemosporidians in blackbirds remain unclear. This study investigates the rate of USUV and Plasmodium spp. co-infection in found-dead blackbirds (n = 203) from 2016 to 2020 in the Netherlands. Presence of Plasmodium spp. was evaluated by cytology (43/203; 21,2%), histopathology (94/186; 50,5%) and qPCR (179/203; 88,1%). The severity of histological lesions in USUV and Plasmodium spp. co-infected dead blackbirds (121/203; 59,6%) were compared with those in Plasmodium spp. single-infected cases. Additionally, since no knowledge is present on the infection rate on live birds and mosquitoes in the Netherlands, a small group of live blackbirds (n = 12) and selected in the field-collected mosquito pools (n = 96) in 2020 were tested for the presence of Plasmodium spp. The latter was detected in the tested live blackbirds by qPCR (8/10; 80%), and cytology (3/11; 27,3%) and in the mosquito pools by qPCR (18/96; 18,7%). For this study, co-infection between USUV and Plasmodium spp. was observed only in the dead blackbirds. The high Plasmodium spp. presence, associated with lower lesions score, in single infected found dead birds suggest a predominantly smaller pathogenic role as single agent. On the other hand, the higher histological lesion scores observed in USUV and Plasmodium spp. co-infected birds suggests a major pathogenic role for the virus or an increased severity of the lesions due to a possible interplay of the two agents.

Keywords: Arboviruses; Avian malaria; Co-infection; Flaviviruses; Zoonosis.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

**Fig. 1**
Lesion patterns and parasitic elements in various tissues of blackbirds infected with *Plasmodium* spp. A. Blood smear from a live blackbird; An erythrocyte shows a round (about 4–6 μm in diameter) light bluish intracytoplasmic element (gametocyte) (short arrowhead) associated with malarial pigment (hemozoin) (x1000). B—F. Histological section of: B. liver; Focal-extensive area of necrosis (*) associated with inflammatory infiltrates and exoerythrocytic *Plasmodium* spp. stages (x400) represented as intracytoplasmic oval structure, up to 60 μm in diameter, containing numerous round 1 μm basophilic elements (merozoites). C. myocardium; A single intra-histiocytic exoerythrocytic *Plasmodium* spp. stage (long arrowhead) is seen in areas of myocarditis (x400). D. lung; Numerous exoerythrocytic *Plasmodium* spp. stage (phanerozoite) in pulmonary capillaries (long arrowhead) represented as an elongated structure, up to 80–100 × 20 μm, containing numerous round 1 μm in diameter basophilic elements (merozoites), are associated with pulmonary edema (x400). E. spleen; Multifocal necrosis and aggregates of brown granular malarial pigment (hemozoin) (x40). F. brain; exoerythrocytic *Plasmodium* spp. stage (phanerozoite) (long arrowhead) in a cerebral capillary surrounded by rare lymphocytes (x400).

**Fig. 2**
Year-round (2016–2020) presence of *Plasmodium* spp. in e ither the liver or the lung of found dead Eurasian blackbirds.

**Fig. 3**
Lesion severity scores by histopathology of *Plasmodium* spp. single infected and USUV co-infected blackbirds.

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References

1. Benzarti E., Sarlet M., Franssen M., Cadar D., Schmidt-Chanasit J., Rivas J.F., Linden A., Desmecht D., Garigliany M. Usutu virus epizootic in Belgium in 2017 and 2018: evidence of virus Endemization and ongoing introduction events. Vector-Borne Zoonot. Diseas. 2020;20 doi: 10.1089/vbz.2019.2469. - DOI - PubMed
1. Weissenböck H., Kolodziejek J., Fragner K., Kuhn R., Pfeffer M., Nowotny N. Usutu virus activity in Austria, 2001–2002. Microbes Infect. 2003;5:1132–1136. doi: 10.1016/S1286-4579(03)00204-1. - DOI - PubMed
1. Ziegler U., Jöst H., Müller K., Fischer D., Rinder M., Tietze D.T., Danner K.-J., Becker N., Skuballa J., Hamann H.-P., Bosch S., Fast C., Eiden M., Schmidt-Chanasit J., Groschup M.H. Epidemic Spread of Usutu Virus in Southwest Germany in 2011 to 2013 and monitoring of wild birds for Usutu and West Nile viruses. Vector-Borne Zoonot. Diseas. 2015;15:481–488. doi: 10.1089/vbz.2014.1746. - DOI - PubMed
1. Garigliany M., Linden A., Gilliau G., Levy E., Sarlet M., Franssen M., Benzarti E., Derouaux A., Francis F., Desmecht D. Usutu virus, Belgium, 2016, Infection. Genetics Evol. 2017;48:116–119. doi: 10.1016/j.meegid.2016.12.023. - DOI - PubMed
1. Höfle U., Gamino V., Fernández de Mera I.G., Mangold A.J., Ortíz J.A., de la Fuente J. Usutu virus in migratory song thrushes, Spain. Emerg. Infect. Dis. 2013;19:1173–1175. doi: 10.3201/eid1907.130199. - DOI - PMC - PubMed

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[1] Benzarti E., Sarlet M., Franssen M., Cadar D., Schmidt-Chanasit J., Rivas J.F., Linden A., Desmecht D., Garigliany M. Usutu virus epizootic in Belgium in 2017 and 2018: evidence of virus Endemization and ongoing introduction events. Vector-Borne Zoonot. Diseas. 2020;20 doi: 10.1089/vbz.2019.2469. - DOI - PubMed

[2] Benzarti E., Sarlet M., Franssen M., Cadar D., Schmidt-Chanasit J., Rivas J.F., Linden A., Desmecht D., Garigliany M. Usutu virus epizootic in Belgium in 2017 and 2018: evidence of virus Endemization and ongoing introduction events. Vector-Borne Zoonot. Diseas. 2020;20 doi: 10.1089/vbz.2019.2469. - DOI - PubMed

[3] Weissenböck H., Kolodziejek J., Fragner K., Kuhn R., Pfeffer M., Nowotny N. Usutu virus activity in Austria, 2001–2002. Microbes Infect. 2003;5:1132–1136. doi: 10.1016/S1286-4579(03)00204-1. - DOI - PubMed

[4] Weissenböck H., Kolodziejek J., Fragner K., Kuhn R., Pfeffer M., Nowotny N. Usutu virus activity in Austria, 2001–2002. Microbes Infect. 2003;5:1132–1136. doi: 10.1016/S1286-4579(03)00204-1. - DOI - PubMed

[5] Ziegler U., Jöst H., Müller K., Fischer D., Rinder M., Tietze D.T., Danner K.-J., Becker N., Skuballa J., Hamann H.-P., Bosch S., Fast C., Eiden M., Schmidt-Chanasit J., Groschup M.H. Epidemic Spread of Usutu Virus in Southwest Germany in 2011 to 2013 and monitoring of wild birds for Usutu and West Nile viruses. Vector-Borne Zoonot. Diseas. 2015;15:481–488. doi: 10.1089/vbz.2014.1746. - DOI - PubMed

[6] Ziegler U., Jöst H., Müller K., Fischer D., Rinder M., Tietze D.T., Danner K.-J., Becker N., Skuballa J., Hamann H.-P., Bosch S., Fast C., Eiden M., Schmidt-Chanasit J., Groschup M.H. Epidemic Spread of Usutu Virus in Southwest Germany in 2011 to 2013 and monitoring of wild birds for Usutu and West Nile viruses. Vector-Borne Zoonot. Diseas. 2015;15:481–488. doi: 10.1089/vbz.2014.1746. - DOI - PubMed

[7] Garigliany M., Linden A., Gilliau G., Levy E., Sarlet M., Franssen M., Benzarti E., Derouaux A., Francis F., Desmecht D. Usutu virus, Belgium, 2016, Infection. Genetics Evol. 2017;48:116–119. doi: 10.1016/j.meegid.2016.12.023. - DOI - PubMed

[8] Garigliany M., Linden A., Gilliau G., Levy E., Sarlet M., Franssen M., Benzarti E., Derouaux A., Francis F., Desmecht D. Usutu virus, Belgium, 2016, Infection. Genetics Evol. 2017;48:116–119. doi: 10.1016/j.meegid.2016.12.023. - DOI - PubMed

[9] Höfle U., Gamino V., Fernández de Mera I.G., Mangold A.J., Ortíz J.A., de la Fuente J. Usutu virus in migratory song thrushes, Spain. Emerg. Infect. Dis. 2013;19:1173–1175. doi: 10.3201/eid1907.130199. - DOI - PMC - PubMed

[10] Höfle U., Gamino V., Fernández de Mera I.G., Mangold A.J., Ortíz J.A., de la Fuente J. Usutu virus in migratory song thrushes, Spain. Emerg. Infect. Dis. 2013;19:1173–1175. doi: 10.3201/eid1907.130199. - DOI - PMC - PubMed

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The pathology of co-infection with Usutu virus and Plasmodium spp. in naturally infected Eurasian blackbirds (Turdus merula)

Affiliations

The pathology of co-infection with Usutu virus and Plasmodium spp. in naturally infected Eurasian blackbirds (Turdus merula)

Authors

Affiliations

Abstract

Conflict of interest statement

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