# Highly Diverse Arenaviruses in Neotropical Bats, Brazil

Luiz Gustavo Bentim Góes, Carlo Fischer, Angélica Cristine Almeida Campos, Cristiano de Carvalho, Andrés Moreira-Soto, Guilherme Ambar, Adriana Ruckert da Rosa, Debora Cardoso de Oliveira, Wendy Karen Jo, Ariovaldo Cruz-Neto, et al.

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## To cite this version:

Luiz Gustavo Bentim Góes, Carlo Fischer, Angélica Cristine Almeida Campos, Cristiano de Carvalho, Andrés Moreira-Soto, et al. Highly Diverse Arenaviruses in Neotropical Bats, Brazil. Emerging Infectious Diseases, 2022, 28 (12), pp.2528-2533. 10.3201/eid2812.220980. hal-04627410

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# Highly Diverse Arenaviruses in Neotropical Bats, Brazil

Luiz Gustavo Bentim Góes, Carlo Fischer, Angélica Cristine Almeida Campos, Cristiano de Carvalho, Andrés Moreira-Soto, Guilherme Ambar, Adriana Ruckert da Rosa, Debora Cardoso de Oliveira, Wendy Karen Jo, Ariovaldo P. Cruz-Neto, Wagner André Pedro, Luzia Helena Queiroz, Paola Minoprio, Edison L. Durigon, Jan Felix Drexler

We detected arenavirus RNA in 1.6% of 1,047 bats in Brazil that were sampled during 2007–2011. We identified Tacaribe virus in 2 *Artibeus* sp. bats and a new arenavirus species in *Carollia perspicillata* bats that we named *Tietê mammarenavirus*. Our results suggest that bats are an underrecognized arenavirus reservoir.

Bats are prominent hosts of zoonotic RNA viruses because of immunologic, physiologic, and ecologic factors (1). The Arenaviridae family comprises 4 genera: *Reptarenavirus* and *Hartmanivirus*, whose members infect reptiles; *Antennavirus*, whose members infect fish; and *Mammarenavirus*, whose members infect mammals. Mammarennaviruses can be separated into globally distributed lymphocytic choriomeningitis–Lassa virus serocomplex and New World arenaviruses (NWAs) (2). The NWAs Junin, Machupo, Sabia, Chapare, and Guanarito cause viral hemorrhagic fever and must be handled under Bio-safety Level 4 conditions (2).

All highly pathogenic arenaviruses known thus far are hosted by and transmitted to humans from persistently infected rodents (2). Only Tacaribe virus (TCRV; *Tacaribe mammarenavirus*) has been identified in bats (3,4). Although TCRV is not

considered a human pathogen, anecdotal evidence exists for potential laboratory acquired infection that causes influenza-like symptoms (5,6). In addition, TCRV is phylogenetically related to pathogenic arenaviruses that cause viral hemorrhagic fever; viral properties associated with severe disease, such as evasion of immune responses and cellular tropism, might be conserved in TCRV and genetically related animal arenaviruses (7).

Associations between TCRV and *Artibeus* spp. bats are supported only by limited epidemiologic data, including a single virus isolation and serologic evidence (3,4), considerable illness of bats during experimental infection (5), and isolation of TCRV from mosquitoes and ticks that primarily feed on rodents and rarely on bats (3,6). Limited genetic data exist for TCRV; a single genomic sequence was obtained from a bat-derived isolate generated in the 1950s from Trinidad that has been extensively passaged in mice and cell cultures and another from a recent tick-derived isolate (3,4,8).

## The Study

We investigated diverse specimens from 1,047 adult bats belonging to 32 species collected from southeastern Brazil (Appendix, https://wwwnc.cdc.gov/EID/article/28/12/22-0980-App1.pdf). We analyzed a total of 3,670 different tissue specimens, including spleens (n = 893), lungs (n = 889), intestines (n = 973), and livers (n = 915), for arenavirus RNA by using reverse transcription PCR (RT-PCR) (9) modified to promote NWA amplification (Appendix Table 1, Figure 1).

We detected arenavirus RNA in 4 *Artibeus litu-ratus*, 1 *A. planirostris*, and 12 *Carollia perspicillata* bats; the overall detection rate was 1.62% (95% CI 0.95%–2.59%). Arenavirus-positive bats were collected during 2007–2011 from 3 sampling sites located in both forest and urban areas within a 60-km radius (Figure 1), suggesting arenavirus

Author affiliations: Charité-Universitätsmedizin Berlin, Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany (L.G.B. Góes, C. Fischer, A.C.A. Campos, A. Moreira-Soto; W.K. Jo, J.F. Drexler); Scientific Platform Pasteur-USP, São Paulo, Brazil (L.G.B. Góes, A.C.A. Campos, P. Minoprio); Universidade de São Paulo, São Paulo, Brazil (L.G.B. Góes, A.C.A. Campos, E.L. Durigon); Universidade Estadual Paulista, Araçatuba, Brazil (C. de Carvalho, W.A. Pedro, L.H. Queiroz); Universidade Estadual Paulista, Rio Claro, Brazil (G. Ambar, A.P. Cruz-Neto), Centro de Controle de Zoonooses, São Paulo (A. Ruckert da Rosa, D. Cardoso de Oliveira); German Centre for Infection Research (DZIF), Berlin (W.K. Jo, J.F. Drexler)

DOI: https://doi.org/10.3201/eid2812.220980
# Highly Diverse Arenaviruses in Neotropical Bats, Brazil

Luiz Gustavo Bentim Góes, Carlo Fischer, Angélica Cristine Almeida Campos, Cristiano de Carvalho, Andrés Moreira-Soto, Guilherme Ambar, Adriana Ruckert da Rosa, Debora Cardoso de Oliveira, Wendy Karen Jo, Ariovaldo P. Cruz-Neto, Wagner André Pedro, Luzia Helena Queiroz, Paola Minoprio, Edison L. Durigon, Jan Felix Drexler

We detected arenavirus RNA in 1.6% of 1,047 bats in Brazil that were sampled during 2007–2011. We identified Tacaribe virus in 2 *Artibeus* sp. bats and a new arenavirus species in *Carollia perspicillata* bats that we named *Tietê mammarenavirus*. Our results suggest that bats are an underrecognized arenavirus reservoir.

Bats are prominent hosts of zoonotic RNA viruses because of immunologic, physiologic, and ecologic factors (1). The Arenaviridae family comprises 4 genera: *Reptarenavirus* and *Hartmanivirus*, whose members infect reptiles; *Antennavirus*, whose members infect fish; and *Mammarenavirus*, whose members infect mammals. Mammarennaviruses can be separated into globally distributed lymphocytic choriomeningitis–Lassa virus serocomplex and New World arenaviruses (NWAs) (2). The NWAs Junin, Machupo, Sabia, Chapare, and Guanarito cause viral hemorrhagic fever and must be handled under Bio-safety Level 4 conditions (2).

All highly pathogenic arenaviruses known thus far are hosted by and transmitted to humans from persistently infected rodents (2). Only Tacaribe virus (TCRV; *Tacaribe mammarenavirus*) has been identified in bats (3,4). Although TCRV is not

considered a human pathogen, anecdotal evidence exists for potential laboratory acquired infection that causes influenza-like symptoms (5,6). In addition, TCRV is phylogenetically related to pathogenic arenaviruses that cause viral hemorrhagic fever; viral properties associated with severe disease, such as evasion of immune responses and cellular tropism, might be conserved in TCRV and genetically related animal arenaviruses (7).

Associations between TCRV and *Artibeus* spp. bats are supported only by limited epidemiologic data, including a single virus isolation and serologic evidence (3,4), considerable illness of bats during experimental infection (5), and isolation of TCRV from mosquitoes and ticks that primarily feed on rodents and rarely on bats (3,6). Limited genetic data exist for TCRV; a single genomic sequence was obtained from a bat-derived isolate generated in the 1950s from Trinidad that has been extensively passaged in mice and cell cultures and another from a recent tick-derived isolate (3,4,8).

## The Study

We investigated diverse specimens from 1,047 adult bats belonging to 32 species collected from southeastern Brazil (Appendix, https://wwwnc.cdc.gov/EID/article/28/12/22-0980-App1.pdf). We analyzed a total of 3,670 different tissue specimens, including spleens (n = 893), lungs (n = 889), intestines (n = 973), and livers (n = 915), for arenavirus RNA by using reverse transcription PCR (RT-PCR) (9) modified to promote NWA amplification (Appendix Table 1, Figure 1).

We detected arenavirus RNA in 4 *Artibeus litu-ratus*, 1 *A. planirostris*, and 12 *Carollia perspicillata* bats; the overall detection rate was 1.62% (95% CI 0.95%–2.59%). Arenavirus-positive bats were collected during 2007–2011 from 3 sampling sites located in both forest and urban areas within a 60-km radius (Figure 1), suggesting arenavirus

Author affiliations: Charité-Universitätsmedizin Berlin, Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany (L.G.B. Góes, C. Fischer, A.C.A. Campos, A. Moreira-Soto; W.K. Jo, J.F. Drexler); Scientific Platform Pasteur-USP, São Paulo, Brazil (L.G.B. Góes, A.C.A. Campos, P. Minoprio); Universidade de São Paulo, São Paulo, Brazil (L.G.B. Góes, A.C.A. Campos, E.L. Durigon); Universidade Estadual Paulista, Araçatuba, Brazil (C. de Carvalho, W.A. Pedro, L.H. Queiroz); Universidade Estadual Paulista, Rio Claro, Brazil (G. Ambar, A.P. Cruz-Neto), Centro de Controle de Zoonooses, São Paulo (A. Ruckert da Rosa, D. Cardoso de Oliveira); German Centre for Infection Research (DZIF), Berlin (W.K. Jo, J.F. Drexler)

DOI: https://doi.org/10.3201/eid2812.220980
<figure><img src="image_1.png" /><figcaption>**Figure 1.** Bat mammarenavirus detection and host distribution in study of highly diverse arenaviruses in neotropical bats, Brazil. A) Geographic ranges of arenavirus-positive bat species indicated by blue (*Artibeus planirostris*), green (*A. lituratus*), and red (*Carollia perspicillata*) colors, according to the International Union for Conservation of Nature (https://www.iucnredlist.org). The brown areas in the map indicate the overlap of the distribution of *A. lituratus* and *C. perspicillata*. The absence of *A. planirostris* distribution in central Brazil likely represents lack of information regarding this species. Filled circles represent regions of sample collection: northwestern region of São Paulo state (red), central region of Paraná state (pink), National Park of Iguaçu, Paraná state (dark blue), and southwestern region of São Paulo state (gray). Number of bats obtained from each region is indicated. Red bat figure indicates where *Tacaribe mammarenavirus* and *Tietê mammarenavirus* were detected in the present study. Hosts from which Tacaribe virus was sequenced in other studies, including ticks (Florida, USA), mosquitoes, and bats (Port of Spain, Trinidad and Tobago) are indicated by black pictograms. Map prepared using QGIS desktop software version 3.24 (https://www.qgis.org). B) Areas of arenavirus detection in the northwestern region of São Paulo state, Brazil. Yellow star indicates the capture site of arenavirus-positive *A. planirostris*, blue star indicates the capture site of arenavirus-positive *A. lituratus*, and orange star marks the capture site of arenavirus-positive *C. perspicillata* bats. Tietê River and cities Araçatuba, Valparaíso, and Birigui are indicated. Dark green areas show forest fragments. Map obtained from Google Earth (https://earth.google.com)</figcaption></figure>

maintenance in bat populations in this region. All 3 arenavirus-positive bat species are abundant in tropical environments and well-adapted to urban landscapes, indicating potential for dispersion and spillover to humans and other animals.

Most arenavirus-positive bats were collected in 2 forest fragments in 2007 (Tables 1, 2; Figure 1), where most bat species positive for arenavirus RNA were sampled. Whether high detection rates at those sites correspond to epizootics or sampling bias remains unknown.

All arenavirus-positive animals appeared healthy, suggesting limited negative effects of arenavirus infection on bat hosts. This observation was similar in rodent arenavirus hosts (10) and consistent with high TCRV seroprevalence in a serologic survey

(4) but different from experimental TCRV infections (5), likely because of different routes and high doses used for infecting bats in laboratory settings. High seroprevalence and low arenavirus detection rates suggest that arenaviruses do not infect bats persistently, which is distinct from results for rodent arenavirus infections (11). Lack of persistence is important for public health because it indicates potential limitations of arenavirus shedding by bat hosts whose lifespan is ≤8-12 years (12).

We detected arenavirus RNA in multiple organs at similar concentrations, including spleens (mean, 1.2 × 10⁷ RNA copies/mg) and lungs (mean, 6.4 × 10⁶ RNA copies/mg) (p = 0.53 by Mann-Whitney U test) (Table 2), suggesting systemic infection similar to that observed in experimentally infected bats (5). We
<figure><img src="image_3.png" /></figure>

**Figure 1.** Bat mammarenavirus detection and host distribution in study of highly diverse arenaviruses in neotropical bats, Brazil. A) Geographic ranges of arenavirus-positive bat species indicated by blue (*Artibeus planirostris*), green (*A. lituratus*), and red (*Carollia perspicillata*) colors, according to the International Union for Conservation of Nature (https://www.iucnredlist.org). The brown areas in the map indicate the overlap of the distribution of *A. lituratus* and *C. perspicillata*. The absence of *A. planirostris* distribution in central Brazil likely represents lack of information regarding this species. Filled circles represent regions of sample collection: northwestern region of São Paulo state (red), central region of Paraná state (pink), National Park of Iguaçu, Paraná state (dark blue), and southwestern region of São Paulo state (gray). Number of bats obtained from each region is indicated. Red bat figure indicates where *Tacaribe mammarenavirus* and *Tietê mammarenavirus* were detected in the present study. Hosts from which Tacaribe virus was sequenced in other studies, including ticks (Florida, USA), mosquitoes, and bats (Port of Spain, Trinidad and Tobago) are indicated by black pictograms. Map prepared using QGIS desktop software version 3.24 (https://www.qgis.org). B) Areas of arenavirus detection in the northwestern region of São Paulo state, Brazil. Yellow star indicates the capture site of arenavirus-positive *A. planirostris*, blue star indicates the capture site of arenavirus-positive *A. lituratus*, and orange star marks the capture site of arenavirus-positive *C. perspicillata* bats. Tietê River and cities Araçatuba, Valparaíso, and Birigui are indicated. Dark green areas show forest fragments. Map obtained from Google Earth (https://earth.google.com)

maintenance in bat populations in this region. All 3 arenavirus-positive bat species are abundant in tropical environments and well-adapted to urban landscapes, indicating potential for dispersion and spillover to humans and other animals.

Most arenavirus-positive bats were collected in 2 forest fragments in 2007 (Tables 1, 2; Figure 1), where most bat species positive for arenavirus RNA were sampled. Whether high detection rates at those sites correspond to epizootics or sampling bias remains unknown.

All arenavirus-positive animals appeared healthy, suggesting limited negative effects of arenavirus infection on bat hosts. This observation was similar in rodent arenavirus hosts (10) and consistent with high TCRV seroprevalence in a serologic survey

(4) but different from experimental TCRV infections (5), likely because of different routes and high doses used for infecting bats in laboratory settings. High seroprevalence and low arenavirus detection rates suggest that arenaviruses do not infect bats persistently, which is distinct from results for rodent arenavirus infections (11). Lack of persistence is important for public health because it indicates potential limitations of arenavirus shedding by bat hosts whose lifespan is ≤8-12 years (12).

We detected arenavirus RNA in multiple organs at similar concentrations, including spleens (mean, 1.2 × 10⁷ RNA copies/mg) and lungs (mean, 6.4 × 10⁶ RNA copies/mg) (p = 0.53 by Mann-Whitney U test) (Table 2), suggesting systemic infection similar to that observed in experimentally infected bats (5). We
observed the highest arenavirus RNA concentration in the single arenavirus-positive intestine specimen, followed by the spleen, lung, liver, and kidney in that animal (Table 2). High arenavirus RNA concentrations in intestines are consistent with virus shedding through the enteric route, which has been observed during experimental infections with TCRV (5). Although rodents shed arenaviruses primarily through urine and saliva, shedding also occurs in feces (2). Determining differences in arenavirus transmission routes between bats and rodents will require further investigation. We were unsuccessful isolating bat arenaviruses from organ homogenates despite repeated attempts (Appendix), likely because of tissue degradation under tropical conditions.

We performed phylogenetic analysis of the partial sequence for the arenavirus RNA-dependent RNA polymerase gene obtained from RT-PCR screening. We found 2 NWA clades in bats from Brazil: 1 clade for both *Artibeus* spp. and 1 clade for *C. perspicillata* bats (GenBank accession nos. ON648806-16) (Figure 2, panel A). We obtained complete arenavirus coding sequences from 1 *A. planirostris* and 3 *C. perspicillata* bats (GenBank accession nos. ON648817-24) by using Illumina-based deep sequencing (Illumina, https://www.illumina.com); genome organization was identical to other mammarennaviruses. Both arenaviruses formed a well-supported monophyletic clade with TCRV in sister relationship to Junin and Machupo viruses (Figure 2, panel B) and

<table><caption>**Table 1.** Bat species screened for arenaviruses in study of highly diverse arenaviruses in neotropical bats, Brazil</caption><thead><tr><th>Bat species</th><th>Family</th><th>No. bats</th><th>No. positive (%, 95% CI)*</th><th>Region†</th><th>Sampling year (no. bats)</th></tr></thead><tbody><tr><td>Artibeus fimbriatus</td><td>Phyllostomidae</td><td>3</td><td>0</td><td>A</td><td>2012 (3)</td></tr><tr><td>A. lituratus</td><td>Phyllostomidae</td><td>155</td><td>4 (2.6, 0.7-6.5)</td><td>A-D</td><td>2007 (8), 2010 (26), 2011 (46), 2012 (45), 2013 (4), 2014 (12), 2015 (16)</td></tr><tr><td>A. obscurus</td><td>Phyllostomidae</td><td>2</td><td>0</td><td>C, D</td><td>2013, 2015</td></tr><tr><td>A. planirostris</td><td>Phyllostomidae</td><td>9</td><td>1 (11.1, 0.3-48.3)</td><td>A-C</td><td>2010 (3), 2011 (2), 2012 (2), 2013 (1), 2014 (1)</td></tr><tr><td>Carollia perspicillata</td><td>Phyllostomidae</td><td>63</td><td>12 (19.1, 10.3-30.9)</td><td>A-D</td><td>2007 (18), 2010 (13), 2011 (18), 2012 (12), 2015 (2)</td></tr><tr><td>Chrotopterus auritus</td><td>Phyllostomidae</td><td>1</td><td>0</td><td>A</td><td>2010</td></tr><tr><td>Cynomops planirostris</td><td>Molossidae</td><td>11</td><td>0</td><td>C, D</td><td>2013 (1), 2014 (7), 2015 (3)</td></tr><tr><td>Desmodus rotundus</td><td>Phyllostomidae</td><td>69</td><td>0</td><td>C, D</td><td>2007 (7), 2011 (44), 2012 (1), 2014 (15), 2015 (2)</td></tr><tr><td>Eptesicus furinalis</td><td>Vespertilionidae</td><td>17</td><td>0</td><td>C, D</td><td>2011 (2), 2013 (6), 2014 (3), 2015 (6)</td></tr><tr><td>Eumops auripendulus</td><td>Molossidae</td><td>2</td><td>0</td><td>D</td><td>2014, 2015</td></tr><tr><td>E. glaucinus</td><td>Molossidae</td><td>106</td><td>0</td><td>C, D</td><td>2009 (1), 2010 (1), 2011 (5), 2012 (5), 2013 (19), 2014 (34), 2015 (41)</td></tr><tr><td>E. perotis</td><td>Molossidae</td><td>12</td><td>0</td><td>C, D</td><td>2013 (1), 2014 (8), 2015 (3)</td></tr><tr><td>Glossophaga soricina</td><td>Phyllostomidae</td><td>70</td><td>0</td><td>C, D</td><td>2007 (3), 2011 (2), 2012 (1), 2013 (2), 2014 (30), 2015 (32)</td></tr><tr><td>Lasiorus blossevillii</td><td>Vespertilionidae</td><td>2</td><td>0</td><td>C, D</td><td>2011, 2012</td></tr><tr><td>L. cinereus</td><td>Vespertilionidae</td><td>1</td><td>0</td><td>C</td><td>2013</td></tr><tr><td>L. ega</td><td>Vespertilionidae</td><td>2</td><td>0</td><td>C, D</td><td>2013, 2014</td></tr><tr><td>Molossops neglectus</td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014</td></tr><tr><td>M. temminckii</td><td>Molossidae</td><td>2</td><td>0</td><td>C</td><td>2011</td></tr><tr><td>Molossus molossus</td><td>Molossidae</td><td>242</td><td>0</td><td>C, D</td><td>2007 (1), 2010 (1), 2011 (25), 2012 (16), 2013 (60), 2014 (84), 2015 (55)</td></tr><tr><td>M. rufus</td><td>Molossidae</td><td>160</td><td>0</td><td>C, D</td><td>2009 (11), 2010 (1), 2011 (20), 2012 (28), 2013 (48), 2014 (27), 2015 (25)</td></tr><tr><td>Myotis nigricans</td><td>Vespertilionidae</td><td>35</td><td>0</td><td>C, D</td><td>2011 (1), 2012 (4), 2013 (9), 2014 (6), 2015 (15)</td></tr><tr><td>M. riparius</td><td>Vespertilionidae</td><td>1</td><td>0</td><td>C</td><td>2013</td></tr><tr><td>Noctilio albiventris</td><td>Noctilionidae</td><td>2</td><td>0</td><td>C</td><td>2007 (2)</td></tr><tr><td>Nyctinomops laticaudatus</td><td>Molossidae</td><td>4</td><td>0</td><td>C, D</td><td>2011 (1), 2014 (2), 2015 (1)</td></tr><tr><td>N. macrotis</td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014 (1)</td></tr><tr><td>Phyllostomus discolor</td><td>Phyllostomidae</td><td>2</td><td>0</td><td>D</td><td>2014 (2)</td></tr><tr><td>Platyrhinus lineatus</td><td>Phyllostomidae</td><td>6</td><td>0</td><td>C, D</td><td>2014 (5), 2015 (1)</td></tr><tr><td>Promops nasutus</td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014 (1)</td></tr><tr><td>Pygoderma bilabiatum</td><td>Phyllostomidae</td><td>1</td><td>0</td><td>D</td><td>2015 (1)</td></tr><tr><td>Sturnira lilium</td><td>Phyllostomidae</td><td>29</td><td>0</td><td>A, B, D</td><td>2010 (5), 2011 (9), 2012 (14), 2015 (1)</td></tr><tr><td>Tadarida brasiliensis</td><td>Molossidae</td><td>30</td><td>0</td><td>C, D</td><td>2014 (15), 2015 (15)</td></tr><tr><td>Vampyressa pusila</td><td>Phyllostomidae</td><td>1</td><td>0</td><td>B</td><td>2012 (1)</td></tr><tr><td>Not identified</td><td>Phyllostomidae</td><td>4</td><td>0</td><td>C, D</td><td>2011(1), 2013 (2), 2014 (1)</td></tr><tr><td>Total</td><td></td><td>4</td><td>1,047</td><td>A-D</td><td>2007-2015</td></tr></tbody></table>

<footnote>*Number of bats with arenavirus RNA detected by PCR.</footnote>

<footnote>†Bats were collected from 36 sites within 4 main geographic regions of Brazil: A, Iguaçu National Park; B, central region of Parana state; C, northwest São Paulo state; and D, southwest São Paulo state.</footnote>
observed the highest arenavirus RNA concentration in the single arenavirus-positive intestine specimen, followed by the spleen, lung, liver, and kidney in that animal (Table 2). High arenavirus RNA concentrations in intestines are consistent with virus shedding through the enteric route, which has been observed during experimental infections with TCRV (5). Although rodents shed arenaviruses primarily through urine and saliva, shedding also occurs in feces (2). Determining differences in arenavirus transmission routes between bats and rodents will require further investigation. We were unsuccessful isolating bat arenaviruses from organ homogenates despite repeated attempts (Appendix), likely because of tissue degradation under tropical conditions.

We performed phylogenetic analysis of the partial sequence for the arenavirus RNA-dependent RNA polymerase gene obtained from RT-PCR screening. We found 2 NWA clades in bats from Brazil: 1 clade for both *Artibeus* spp. and 1 clade for *C. perspicillata* bats (GenBank accession nos. ON648806-16) (Figure 2, panel A). We obtained complete arenavirus coding sequences from 1 *A. planirostris* and 3 *C. perspicillata* bats (GenBank accession nos. ON648817-24) by using Illumina-based deep sequencing (Illumina, https://www.illumina.com); genome organization was identical to other mammarennaviruses. Both arenaviruses formed a well-supported monophyletic clade with TCRV in sister relationship to Junin and Machupo viruses (Figure 2, panel B) and

<table><caption>**Table 1.** Bat species screened for arenaviruses in study of highly diverse arenaviruses in neotropical bats, Brazil</caption><thead><tr><th>Bat species</th><th>Family</th><th>No. bats</th><th>No. positive (%, 95% CI)*</th><th>Region†</th><th>Sampling year (no. bats)</th></tr></thead><tbody><tr><td><i>Artibeus fimbriatus</i></td><td>Phyllostomidae</td><td>3</td><td>0</td><td>A</td><td>2012 (3)</td></tr><tr><td><i>A. lituratus</i></td><td>Phyllostomidae</td><td>155</td><td>4 (2.6, 0.7-6.5)</td><td>A-D</td><td>2007 (8), 2010 (26), 2011 (46), 2012 (45), 2013 (4), 2014 (12), 2015 (16)</td></tr><tr><td><i>A. obscurus</i></td><td>Phyllostomidae</td><td>2</td><td>0</td><td>C, D</td><td>2013, 2015</td></tr><tr><td><i>A. planirostris</i></td><td>Phyllostomidae</td><td>9</td><td>1 (11.1, 0.3-48.3)</td><td>A-C</td><td>2010 (3), 2011 (2), 2012 (2), 2013 (1), 2014 (1)</td></tr><tr><td><i>Carollia perspicillata</i></td><td>Phyllostomidae</td><td>63</td><td>12 (19.1, 10.3-30.9)</td><td>A-D</td><td>2007 (18), 2010 (13), 2011 (18), 2012 (12), 2015 (2)</td></tr><tr><td><i>Chrotopterus auritus</i></td><td>Phyllostomidae</td><td>1</td><td>0</td><td>A</td><td>2010</td></tr><tr><td><i>Cynomops planirostris</i></td><td>Molossidae</td><td>11</td><td>0</td><td>C, D</td><td>2013 (1), 2014 (7), 2015 (3)</td></tr><tr><td><i>Desmodus rotundus</i></td><td>Phyllostomidae</td><td>69</td><td>0</td><td>C, D</td><td>2007 (7), 2011 (44), 2012 (1), 2014 (15), 2015 (2)</td></tr><tr><td><i>Eptesicus furinalis</i></td><td>Vespertilionidae</td><td>17</td><td>0</td><td>C, D</td><td>2011 (2), 2013 (6), 2014 (3), 2015 (6)</td></tr><tr><td><i>Eumops auripendulus</i></td><td>Molossidae</td><td>2</td><td>0</td><td>D</td><td>2014, 2015</td></tr><tr><td><i>E. glaucinus</i></td><td>Molossidae</td><td>106</td><td>0</td><td>C, D</td><td>2009 (1), 2010 (1), 2011 (5), 2012 (5), 2013 (19), 2014 (34), 2015 (41)</td></tr><tr><td><i>E. perotis</i></td><td>Molossidae</td><td>12</td><td>0</td><td>C, D</td><td>2013 (1), 2014 (8), 2015 (3)</td></tr><tr><td><i>Glossophaga soricina</i></td><td>Phyllostomidae</td><td>70</td><td>0</td><td>C, D</td><td>2007 (3), 2011 (2), 2012 (1), 2013 (2), 2014 (30), 2015 (32)</td></tr><tr><td><i>Lasiorus blossevillii</i></td><td>Vespertilionidae</td><td>2</td><td>0</td><td>C, D</td><td>2011, 2012</td></tr><tr><td><i>L. cinereus</i></td><td>Vespertilionidae</td><td>1</td><td>0</td><td>C</td><td>2013</td></tr><tr><td><i>L. ega</i></td><td>Vespertilionidae</td><td>2</td><td>0</td><td>C, D</td><td>2013, 2014</td></tr><tr><td><i>Molossops neglectus</i></td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014</td></tr><tr><td><i>M. temminckii</i></td><td>Molossidae</td><td>2</td><td>0</td><td>C</td><td>2011</td></tr><tr><td><i>Molossus molossus</i></td><td>Molossidae</td><td>242</td><td>0</td><td>C, D</td><td>2007 (1), 2010 (1), 2011 (25), 2012 (16), 2013 (60), 2014 (84), 2015 (55)</td></tr><tr><td><i>M. rufus</i></td><td>Molossidae</td><td>160</td><td>0</td><td>C, D</td><td>2009 (11), 2010 (1), 2011 (20), 2012 (28), 2013 (48), 2014 (27), 2015 (25)</td></tr><tr><td><i>Myotis nigricans</i></td><td>Vespertilionidae</td><td>35</td><td>0</td><td>C, D</td><td>2011 (1), 2012 (4), 2013 (9), 2014 (6), 2015 (15)</td></tr><tr><td><i>M. riparius</i></td><td>Vespertilionidae</td><td>1</td><td>0</td><td>C</td><td>2013</td></tr><tr><td><i>Noctilio albiventris</i></td><td>Noctilionidae</td><td>2</td><td>0</td><td>C</td><td>2007 (2)</td></tr><tr><td><i>Nyctinomops laticaudatus</i></td><td>Molossidae</td><td>4</td><td>0</td><td>C, D</td><td>2011 (1), 2014 (2), 2015 (1)</td></tr><tr><td><i>N. macrotis</i></td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014 (1)</td></tr><tr><td><i>Phyllostomus discolor</i></td><td>Phyllostomidae</td><td>2</td><td>0</td><td>D</td><td>2014 (2)</td></tr><tr><td><i>Platyrhinus lineatus</i></td><td>Phyllostomidae</td><td>6</td><td>0</td><td>C, D</td><td>2014 (5), 2015 (1)</td></tr><tr><td><i>Promops nasutus</i></td><td>Molossidae</td><td>1</td><td>0</td><td>D</td><td>2014 (1)</td></tr><tr><td><i>Pygoderma bilabiatum</i></td><td>Phyllostomidae</td><td>1</td><td>0</td><td>D</td><td>2015 (1)</td></tr><tr><td><i>Sturnira lilium</i></td><td>Phyllostomidae</td><td>29</td><td>0</td><td>A, B, D</td><td>2010 (5), 2011 (9), 2012 (14), 2015 (1)</td></tr><tr><td><i>Tadarida brasiliensis</i></td><td>Molossidae</td><td>30</td><td>0</td><td>C, D</td><td>2014 (15), 2015 (15)</td></tr><tr><td><i>Vampyressa pusila</i></td><td>Phyllostomidae</td><td>1</td><td>0</td><td>B</td><td>2012 (1)</td></tr><tr><td>Not identified</td><td>Phyllostomidae</td><td>4</td><td>0</td><td>C, D</td><td>2011(1), 2013 (2), 2014 (1)</td></tr><tr><td>Total</td><td></td><td>4</td><td>1,047</td><td>A-D</td><td>2007-2015</td></tr></tbody></table>

<footnote>*Number of bats with arenavirus RNA detected by PCR.</footnote>

<footnote>†Bats were collected from 36 sites within 4 main geographic regions of Brazil: A, Iguaçu National Park; B, central region of Parana state; C, northwest São Paulo state; and D, southwest São Paulo state.</footnote>
<table><caption>**Table 2.** Collection sites and arenavirus RNA concentrations in different organs from bats in study of highly diverse arenaviruses in neotropical bats, Brazil*</caption><thead><tr><th rowspan="2">Sample no.</th><th rowspan="2">Bat species†</th><th rowspan="2">Sex</th><th rowspan="2">Collection site</th><th colspan="5">No. RNA copies/mg tissue</th></tr><tr><th>Spleen</th><th>Lung</th><th>Intestine</th><th>Liver</th><th>Kidney</th></tr></thead><tbody><tr><td>Br56</td><td>Artibeus lituratus</td><td>M</td><td>Valparaiso</td><td>5.54 x 10<sup>2</sup></td><td>7.24 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br57</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>NA</td><td>3.14 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br58</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>2.15 x 10<sup>6</sup></td><td>6.13 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br59</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>NA</td><td>2.10 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>A354</td><td>A. planirostris</td><td>M</td><td>Birigui</td><td>1.09 x 10<sup>5</sup></td><td>5.02 x 10<sup>4</sup></td><td>4.73 x 10<sup>5</sup></td><td>9.68 x 10<sup>3</sup></td><td>6.01 x 10<sup>3</sup></td></tr><tr><td>Br61</td><td>Carollia perspicillata</td><td>F</td><td>Araçatuba</td><td>4.73 x 10<sup>4</sup></td><td>1.17 x 10<sup>3</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br62</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>1.99 x 10<sup>7</sup></td><td>6.96 x 10<sup>7</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br63</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>2.71 x 10<sup>2</sup></td><td>8.61 x 10<sup>°</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br65</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>2.23 x 10<sup>1</sup></td><td>2.88 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br68</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>2.35 x 10<sup>1</sup></td><td>Neg</td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br69</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>5.95 x 10<sup>7</sup></td><td>1.99 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br70</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>8.20 x 10<sup>7</sup></td><td>1.85 x 10<sup>5</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br71</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>5.37 x 10<sup>3</sup></td><td>5.38 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br72</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>4.70 x 10<sup>2</sup></td><td>6.11 x 10<sup>1</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br74</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>3.54 x 10<sup>5</sup></td><td>8.84 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br76</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>1.18 x 10<sup>6</sup></td><td>1.52 x 10<sup>7</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br77</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>Neg</td><td>1.81 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr></tbody></table>

*Numbers in bold are samples used in arenavirus isolation attempts. NA, tissue not available; Neg, negative

†Samples were collected from *Artibeus lituratus* bats in forest areas of Valparaiso in 2007, *A. planirostris* bat in an urban area of Birigui in 2011, and *Carollia perspicillata* bats in forest areas of Araçatuba in 2007.

Ocozocoautla de Espinosa virus that was possibly responsible for a hemorrhagic fever outbreak in Mexico (Figure 2, panel C) (13). These results highlight the

genetic relationship of those bat-associated arenaviruses with highly pathogenic NWAs (Appendix Table 2). Identical topology in phylogenetic reconstructions

<figure><img src="image_2.png" /><figcaption>**Figure 2.** Phylogenetic analyses of highly diverse arenaviruses in neotropical bats, Brazil. Maximum-likelihood consensus trees compare partial RNA-dependent RNA polymerase genes (A), complete large (L) segment genes (B), and complete small (S) segment genes (C) from arenaviruses detected in *Artibeus* and *Carollia* spp. bats. Phylogenetic trees were generated using MEGA X software (https://www.megasoftware.net). Bold indicates sequences obtained from this study. Stars indicate regions where arenavirus-positive bat hosts were detected (Figure 1, panel B). Black dots at tree nodes represent bootstrap values ≥75% (1,000 replicates). Green lines indicate clade A new world arenaviruses, red lines indicate clade B new world arenaviruses, blue lines indicate clade C new world arenaviruses, and purple lines indicate recombinant new world arenaviruses (tentative clade D) (14). GenBank sequences used for comparisons and virus abbreviations are provided online (https://wwwnc.cdc.gov/EID/article/28/12/22-0980-F2.htm). Origins of arenaviruses are indicated for each sample: ARG, Argentina; BOL, Bolivia; BRA, Brazil; COL, Colombia; PER, Peru; TRI, Trinidad; USA, United States of America; VEN, Venezuela. Scale bars indicate nucleotide substitutions per site.</figcaption></figure>
<table><caption>**Table 2.** Collection sites and arenavirus RNA concentrations in different organs from bats in study of highly diverse arenaviruses in neotropical bats, Brazil*</caption><thead><tr><th rowspan="2">Sample no.</th><th rowspan="2">Bat species†</th><th rowspan="2">Sex</th><th rowspan="2">Collection site</th><th colspan="5">No. RNA copies/mg tissue</th></tr><tr><th>Spleen</th><th>Lung</th><th>Intestine</th><th>Liver</th><th>Kidney</th></tr></thead><tbody><tr><td>Br56</td><td>Artibeus lituratus</td><td>M</td><td>Valparaiso</td><td>5.54 x 10<sup>2</sup></td><td>7.24 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br57</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>NA</td><td>3.14 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br58</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>2.15 x 10<sup>6</sup></td><td>6.13 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br59</td><td>A. lituratus</td><td>M</td><td>Valparaiso</td><td>NA</td><td>2.10 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>A354</td><td>A. planirostris</td><td>M</td><td>Birigui</td><td>1.09 x 10<sup>5</sup></td><td>5.02 x 10<sup>4</sup></td><td>4.73 x 10<sup>5</sup></td><td>9.68 x 10<sup>3</sup></td><td>6.01 x 10<sup>3</sup></td></tr><tr><td>Br61</td><td>Carollia perspicillata</td><td>F</td><td>Araçatuba</td><td>4.73 x 10<sup>4</sup></td><td>1.17 x 10<sup>3</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br62</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>1.99 x 10<sup>7</sup></td><td>6.96 x 10<sup>7</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br63</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>2.71 x 10<sup>2</sup></td><td>8.61 x 10<sup>°</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br65</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>2.23 x 10<sup>1</sup></td><td>2.88 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br68</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>2.35 x 10<sup>1</sup></td><td>Neg</td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br69</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>5.95 x 10<sup>7</sup></td><td>1.99 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br70</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>8.20 x 10<sup>7</sup></td><td>1.85 x 10<sup>5</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br71</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>5.37 x 10<sup>3</sup></td><td>5.38 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br72</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>4.70 x 10<sup>2</sup></td><td>6.11 x 10<sup>1</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br74</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>3.54 x 10<sup>5</sup></td><td>8.84 x 10<sup>6</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br76</td><td>C. perspicillata</td><td>M</td><td>Araçatuba</td><td>1.18 x 10<sup>6</sup></td><td>1.52 x 10<sup>7</sup></td><td>NA</td><td>NA</td><td>NA</td></tr><tr><td>Br77</td><td>C. perspicillata</td><td>F</td><td>Araçatuba</td><td>Neg</td><td>1.81 x 10<sup>2</sup></td><td>NA</td><td>NA</td><td>NA</td></tr></tbody></table>

*Numbers in bold are samples used in arenavirus isolation attempts. NA, tissue not available; Neg, negative

†Samples were collected from *Artibeus lituratus* bats in forest areas of Valparaiso in 2007, *A. planirostris* bat in an urban area of Birigui in 2011, and *Carollia perspicillata* bats in forest areas of Araçatuba in 2007.

Ocozocoautla de Espinosa virus that was possibly responsible for a hemorrhagic fever outbreak in Mexico (Figure 2, panel C) (13). These results highlight the

genetic relationship of those bat-associated arenaviruses with highly pathogenic NWAs (Appendix Table 2). Identical topology in phylogenetic reconstructions

<figure><img src="image_4.png" /><figcaption>**Figure 2.** Phylogenetic analyses of highly diverse arenaviruses in neotropical bats, Brazil. Maximum-likelihood consensus trees compare partial RNA-dependent RNA polymerase genes (A), complete large (L) segment genes (B), and complete small (S) segment genes (C) from arenaviruses detected in *Artibeus* and *Carollia* spp. bats. Phylogenetic trees were generated using MEGA X software (https://www.megasoftware.net). Bold indicates sequences obtained from this study. Stars indicate regions where arenavirus-positive bat hosts were detected (Figure 1, panel B). Black dots at tree nodes represent bootstrap values ≥75% (1,000 replicates). Green lines indicate clade A new world arenaviruses, red lines indicate clade B new world arenaviruses, blue lines indicate clade C new world arenaviruses, and purple lines indicate recombinant new world arenaviruses (tentative clade D) (14). GenBank sequences used for comparisons and virus abbreviations are provided online (https://wwwnc.cdc.gov/EID/article/28/12/22-0980-F2.htm). Origins of arenaviruses are indicated for each sample: ARG, Argentina; BOL, Bolivia; BRA, Brazil; COL, Colombia; PER, Peru; TRI, Trinidad; USA, United States of America; VEN, Venezuela. Scale bars indicate nucleotide substitutions per site.</figcaption></figure>
argued against potential reassortment (Figure 2, panels B, C), and homogeneous sequence distances and recombination analyses along the genome did not indicate recombination events (Appendix Figure 2).

The *A. planirostris* bat was infected with a previously unknown TCRV strain (Appendix Table 2) that had an amino acid identity of 93.8%–95.5% with other TCRV sequences, depending on the protein analyzed. The arenaviruses from *C. perspicillata* bats formed a separate species in clade B of the TCRV serogroup (Figure 2, panels B, C). Species assignment relied on taxonomic criteria (14) that included exclusive detection in a distinct host, nucleotide sequence identity of <80% in the small segment, and 88.6%–90% amino acid identity in the nucleocapsid protein compared with TCRV and pairwise sequence comparison (https://www.ncbi.nlm.nih.gov/sutils/pasc/viridity.cgi?textpage=overview) results for large and small segments (Appendix Figure 3). The 5' and 3' ends of large and small genomic segments obtained from the newly identified arenavirus from *C. perspicillata* bats were nearly identical to TCRV, consistent with a close genetic relationship between those NWAs (Appendix Table 3, Figure 4). We propose that the arenavirus sequenced from *C. perspicillata* bats should be named Tietê virus (species *Tiête mammarenavirus*) and abbreviated as TETV; the name comes from the main river located <4 km from the capture site (Figure 1).

## Conclusions

Arenavirus genetic diversity is hypothesized to result from a complex macro-evolutionary pattern that includes both co-evolution and host switching in the Muridae family of rodents. In South America, arenaviruses might have co-evolved with rodents in the Sigmodontinae subfamily, with the exception of TCRV (10). Further investigation will be required to determine whether bat arenaviruses evolved from an ancestral host switch involving rodents, which would be consistent with the genetic relationship between TCRV or Tietê virus and rodent-derived Ocozocoautla de Espinosa virus, or whether bats and arenaviruses co-evolved. Of note, bats play an essential role in ecosystems, and stigmatization of bats as sources of zoonotic viruses is unwarranted.

In summary, the epidemiology, genealogy, and zoonotic potential of bat arenaviruses deserve further investigation. Our results suggest that bats are an underrecognized arenavirus reservoir.

Authorization for bat captures was provided by the Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais (approval nos. 12751-2, 12751-3, 21748-1, and

27346-2), Instituto Ambiental do Paraná (approval no. 235/10), ethics committee of the Universidade Estadual Paulista (UNESP) (approval no. 009/2012), and the ethics committee of the Institute of Biomedical Science from the University of São Paulo (approval no. 56-18-03/2014).

This work was supported by the National Council for Scientific and Technological Development–CNPq (grant agreement nos. 152365/2019-2 and 203084/2019-5), São Paulo Research Foundation (FAPESP) (grant agreement nos. 2013/11.006-0 and 2014/15090-8), and Human Fronter Science Program (grant agreement no. RPG0013/2018).

## About the Author

Dr. Góes is a postdoctoral researcher at the Institute of Virology at Charité-Universitätsmedizin, Berlin, Germany. His research focuses on pathogen discovery, virus epidemiology, and evolution of emerging viruses.

## References

1. Guth S, Mollentze N, Renault K, Streicker DG, Visher E, Boots M, et al. Bats host the most virulent—but not the most dangerous—zoonotic viruses. Proc Natl Acad Sci USA. 2022;119:e2113628119. https://doi.org/10.1073/pnas.2113628119

2. Charrel RN, de Lamballerie X. Zoonotic aspects of arenavirus infections. Vet Microbiol. 2010;140:213–20. https://doi.org/10.1016/j.vetmic.2009.08.027

3. Downs WG, Anderson CR, Spence L, Aitken THG, Greenhall AH. Tacaribe virus, a new agent isolated from Artibeus bats and mosquitoes in Trinidad, West Indies. Am J Trop Med Hyg. 1963;12:640–6. https://doi.org/10.4269/ajtmh.1963.12.640

4. Malmlov A, Seetahal J, Carrington C, Ramkisson V, Foster J, Miazgowicz KL, et al. Serological evidence of arenavirus circulation among fruit bats in Trinidad. PLoS One. 2017;12:e0185308. https://doi.org/10.1371/journal.pone.0185308

5. Cogswell-Hawkinson A, Bowen R, James S, Gardiner D, Calisher CH, Adams R, et al. Tacaribe virus causes fatal infection of an ostensible reservoir host, the Jamaican fruit bat. J Virol. 2012;86:5791–9. https://doi.org/10.1128/JVI.00201-12

6. Sayler KA, Barbet AF, Chamberlain C, Clapp WL, Alleman R, Loeb JC, et al. Isolation of Tacaribe virus, a Caribbean arenavirus, from host-seeking *Amblyomma americanum* ticks in Florida. PLoS One. 2014;9:e115769. https://doi.org/10.1371/journal.pone.0115769

7. Moreno H, Möller R, Fedeli C, Gerold G, Kunz S. Comparison of the innate immune responses to pathogenic and nonpathogenic clade B New World arenaviruses. J Virol. 2019;93:e00148-19. https://doi.org/10.1128/JVI.00148-19

8. Holzerland J, Leske A, Fénéant L, Garcin D, Kolakofsky D, Groseth A. Complete genome sequence of Tacaribe virus. Arch Virol. 2020;165:1899–903. https://doi.org/10.1007/s00705-020-04681-9

9. Vieth S, Drosten C, Lenz O, Vincent M, Omilabu S, Hass M, et al. RT-PCR assay for detection of Lassa virus and related Old World arenaviruses targeting the L gene. Trans R Soc
argued against potential reassortment (Figure 2, panels B, C), and homogeneous sequence distances and recombination analyses along the genome did not indicate recombination events (Appendix Figure 2).

The *A. planirostris* bat was infected with a previously unknown TCRV strain (Appendix Table 2) that had an amino acid identity of 93.8%–95.5% with other TCRV sequences, depending on the protein analyzed. The arenaviruses from *C. perspicillata* bats formed a separate species in clade B of the TCRV serogroup (Figure 2, panels B, C). Species assignment relied on taxonomic criteria (14) that included exclusive detection in a distinct host, nucleotide sequence identity of <80% in the small segment, and 88.6%–90% amino acid identity in the nucleocapsid protein compared with TCRV and pairwise sequence comparison (https://www.ncbi.nlm.nih.gov/sutils/pasc/viridity.cgi?textpage=overview) results for large and small segments (Appendix Figure 3). The 5' and 3' ends of large and small genomic segments obtained from the newly identified arenavirus from *C. perspicillata* bats were nearly identical to TCRV, consistent with a close genetic relationship between those NWAs (Appendix Table 3, Figure 4). We propose that the arenavirus sequenced from *C. perspicillata* bats should be named Tietê virus (species *Tiête mammarenavirus*) and abbreviated as TETV; the name comes from the main river located <4 km from the capture site (Figure 1).

## Conclusions

Arenavirus genetic diversity is hypothesized to result from a complex macro-evolutionary pattern that includes both co-evolution and host switching in the Muridae family of rodents. In South America, arenaviruses might have co-evolved with rodents in the Sigmodontinae subfamily, with the exception of TCRV (10). Further investigation will be required to determine whether bat arenaviruses evolved from an ancestral host switch involving rodents, which would be consistent with the genetic relationship between TCRV or Tietê virus and rodent-derived Ocozocoautla de Espinosa virus, or whether bats and arenaviruses co-evolved. Of note, bats play an essential role in ecosystems, and stigmatization of bats as sources of zoonotic viruses is unwarranted.

In summary, the epidemiology, genealogy, and zoonotic potential of bat arenaviruses deserve further investigation. Our results suggest that bats are an underrecognized arenavirus reservoir.

Authorization for bat captures was provided by the Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais (approval nos. 12751-2, 12751-3, 21748-1, and

27346-2), Instituto Ambiental do Paraná (approval no. 235/10), ethics committee of the Universidade Estadual Paulista (UNESP) (approval no. 009/2012), and the ethics committee of the Institute of Biomedical Science from the University of São Paulo (approval no. 56-18-03/2014).

This work was supported by the National Council for Scientific and Technological Development–CNPq (grant agreement nos. 152365/2019-2 and 203084/2019-5), São Paulo Research Foundation (FAPESP) (grant agreement nos. 2013/11.006-0 and 2014/15090-8), and Human Fronter Science Program (grant agreement no. RPG0013/2018).

## About the Author

Dr. Góes is a postdoctoral researcher at the Institute of Virology at Charité-Universitätsmedizin, Berlin, Germany. His research focuses on pathogen discovery, virus epidemiology, and evolution of emerging viruses.

## References

1. Guth S, Mollentze N, Renault K, Streicker DG, Visher E, Boots M, et al. Bats host the most virulent—but not the most dangerous—zoonotic viruses. Proc Natl Acad Sci USA. 2022;119:e2113628119. https://doi.org/10.1073/pnas.2113628119

2. Charrel RN, de Lamballerie X. Zoonotic aspects of arenavirus infections. Vet Microbiol. 2010;140:213–20. https://doi.org/10.1016/j.vetmic.2009.08.027

3. Downs WG, Anderson CR, Spence L, Aitken THG, Greenhall AH. Tacaribe virus, a new agent isolated from Artibeus bats and mosquitoes in Trinidad, West Indies. Am J Trop Med Hyg. 1963;12:640–6. https://doi.org/10.4269/ajtmh.1963.12.640

4. Malmlov A, Seetahal J, Carrington C, Ramkisson V, Foster J, Miazgowicz KL, et al. Serological evidence of arenavirus circulation among fruit bats in Trinidad. PLoS One. 2017;12:e0185308. https://doi.org/10.1371/journal.pone.0185308

5. Cogswell-Hawkinson A, Bowen R, James S, Gardiner D, Calisher CH, Adams R, et al. Tacaribe virus causes fatal infection of an ostensible reservoir host, the Jamaican fruit bat. J Virol. 2012;86:5791–9. https://doi.org/10.1128/JVI.00201-12

6. Sayler KA, Barbet AF, Chamberlain C, Clapp WL, Alleman R, Loeb JC, et al. Isolation of Tacaribe virus, a Caribbean arenavirus, from host-seeking *Amblyomma americanum* ticks in Florida. PLoS One. 2014;9:e115769. https://doi.org/10.1371/journal.pone.0115769

7. Moreno H, Möller R, Fedeli C, Gerold G, Kunz S. Comparison of the innate immune responses to pathogenic and nonpathogenic clade B New World arenaviruses. J Virol. 2019;93:e00148-19. https://doi.org/10.1128/JVI.00148-19

8. Holzerland J, Leske A, Fénéant L, Garcin D, Kolakofsky D, Groseth A. Complete genome sequence of Tacaribe virus. Arch Virol. 2020;165:1899–903. https://doi.org/10.1007/s00705-020-04681-9

9. Vieth S, Drosten C, Lenz O, Vincent M, Omilabu S, Hass M, et al. RT-PCR assay for detection of Lassa virus and related Old World arenaviruses targeting the L gene. Trans R Soc
Trop Med Hyg. 2007;101:1253-64. https://doi.org/10.1016/
j.trstmh.2005.03.018

10. Grande-Pérez A, Martin V, Moreno H, de la Torre JC. Arenavirus quasispecies and their biological implications. Curr Top Microbiol Immunol. 2016;392:231-76. https://doi.org/10.1007/82_2015_468

11. Hoffmann C, Wurr S, Pallasch E, Bockholt S, Rieger T, Günther S, et al. Experimental Morogoro virus infection in its natural host, *Mastomys natalensis*. Viruses. 2021;13:851. https://doi.org/10.3390/v13050851

12. Jones ML. Longevity of captive mammals. Zool Garten N. F. Jena. 1982;52:113-28 [cited 2022 Nov 10]. http://www.rhinoresourcecenter.com/pdf_files/125/1256468598.pdf

13. Cajimat MNB, Milazzo ML, Bradley RD, Fulhorst CF. Ocozocoautla de espinosa virus and hemorrhagic fever, Mexico. Emerg Infect Dis. 2012;18:401-5. https://doi.org/10.3201/eid1803.111602

14. Radoshitzky SR, Bào Y, Buchmeier MJ, Charrel RN, Clawson AN, Clegg CS, et al. Past, present, and future of arenavirus taxonomy. Arch Virol. 2015;160:1851-74. https://doi.org/10.1007/s00705-015-2418-y

Address for correspondence: Jan Felix Drexler, Helmut-Ruska-Haus, Institute of Virology, Campus Charité Mitte, Charitéplatz 1, 10098 Berlin, Germany; email: felix.drexler@charite.de

August 2022
# Zoonotic Infections

* Incidence of Nontuberculous Mycobacterial Pulmonary Infection, by Ethnic Group, Hawaii, USA, 2005–2019

* Investigation of COVID-19 Outbreak among Wildland Firefighters during Wildfire Response, Colorado, USA, 2020

* Lack of Evidence for Ribavirin Treatment of Lassa Fever in Systematic Review of Published and Unpublished Studies

* Dominant Carbapenemase-Encoding Plasmids in Clinical Enterobacterales Isolates and Hypervirulent *Klebsiella pneumoniae*, Singapore

* Increasing and More Commonly Refractory *Mycobacterium avium* Pulmonary Disease, Toronto, Ontario, Canada

* Characterization of Emerging Serotype 19A Pneumococcal Strains in Invasive Disease and Carriage, Belgium

* COVID-19 Symptoms and Deaths among Healthcare Workers, United States

* Association of Environmental Factors with Seasonal Intensity of *Erysipelothrix rhusiopathiae* Seropositivity among Arctic Caribou

EMERGING
INFECTIOUS DISEASES

Zoonotic Infections

CDC

August 2022

<figure><img src="image_3.png" /></figure>

* Factors Associated with Delayed or Missed Second-Dose mRNA COVID-19 Vaccination among Persons >12 Years of Age, United States

* COVID-19 Vaccination Intent and Belief that Vaccination Will End the Pandemic

* Invasive Pneumococcal Disease and Long-Term Mortality Rates in Adults, Alberta, Canada

* Dog Ownership and Risk for Alveolar Echinococcosis, Germany

* Novel Chronic Anaplasmosis in Splenectomized Patient, Amazon Rainforest

* Transmissibility of SARS-CoV-2 B.1.1.214 and Alpha Variants during 4 COVID-19 Waves, Kyoto, Japan, January 2020–June 2021

* Culling of Urban Norway Rats and Carriage of *Bartonella* spp. Bacteria, Vancouver, British Columbia, Canada

* Zoonotic Threat of G4 Genotype Eurasian Avian-Like Swine Influenza A(H1N1) Viruses, China, 2020

* Increased Incidence of Invasive Pneumococcal Disease among Children after COVID-19 Pandemic, England

* Anthelmintic Baiting of Foxes against *Echinococcus multilocularis* in Small Public Area, Japan

* *Spiroplasma ixodetis* Infections in Immunocompetent and Immunosuppressed Patients after Tick Exposure, Sweden

* Child Melioidosis Deaths Caused by *Burkholderia pseudomallei*–Contaminated Borehole Water, Vietnam, 2019

* Association of Phylogenomic Relatedness among *Neisseria gonorrhoeae* Strains with Antimicrobial Resistance, Austria, 2016–2020

EMERGING
INFECTIOUS DISEASES

To revisit the August 2022 issue, go to:
https://wwwnc.cdc.gov/eid/articles/issue/28/8/table-of-contents
Trop Med Hyg. 2007;101:1253-64. https://doi.org/10.1016/
j.trstmh.2005.03.018

10. Grande-Pérez A, Martin V, Moreno H, de la Torre JC. Arenavirus quasispecies and their biological implications. Curr Top Microbiol Immunol. 2016;392:231-76. https://doi.org/10.1007/82_2015_468

11. Hoffmann C, Wurr S, Pallasch E, Bockholt S, Rieger T, Günther S, et al. Experimental Morogoro virus infection in its natural host, *Mastomys natalensis*. Viruses. 2021;13:851. https://doi.org/10.3390/v13050851

12. Jones ML. Longevity of captive mammals. Zool Garten N. F. Jena. 1982;52:113-28 [cited 2022 Nov 10]. http://www.rhinoresourcecenter.com/pdf_files/125/1256468598.pdf

13. Cajimat MNB, Milazzo ML, Bradley RD, Fulhorst CF. Ocozocoautla de espinosa virus and hemorrhagic fever, Mexico. Emerg Infect Dis. 2012;18:401-5. https://doi.org/10.3201/eid1803.111602

14. Radoshitzky SR, Bào Y, Buchmeier MJ, Charrel RN, Clawson AN, Clegg CS, et al. Past, present, and future of arenavirus taxonomy. Arch Virol. 2015;160:1851-74. https://doi.org/10.1007/s00705-015-2418-y

Address for correspondence: Jan Felix Drexler, Helmut-Ruska-Haus, Institute of Virology, Campus Charité Mitte, Charitéplatz 1, 10098 Berlin, Germany; email: felix.drexler@charite.de

August 2022
# Zoonotic Infections

* Incidence of Nontuberculous Mycobacterial Pulmonary Infection, by Ethnic Group, Hawaii, USA, 2005–2019

* Investigation of COVID-19 Outbreak among Wildland Firefighters during Wildfire Response, Colorado, USA, 2020

* Lack of Evidence for Ribavirin Treatment of Lassa Fever in Systematic Review of Published and Unpublished Studies

* Dominant Carbapenemase-Encoding Plasmids in Clinical Enterobacterales Isolates and Hypervirulent *Klebsiella pneumoniae*, Singapore

* Increasing and More Commonly Refractory *Mycobacterium avium* Pulmonary Disease, Toronto, Ontario, Canada

* Characterization of Emerging Serotype 19A Pneumococcal Strains in Invasive Disease and Carriage, Belgium

* COVID-19 Symptoms and Deaths among Healthcare Workers, United States

* Association of Environmental Factors with Seasonal Intensity of *Erysipelothrix rhusiopathiae* Seropositivity among Arctic Caribou

EMERGING
INFECTIOUS DISEASES

Zoonotic Infections

CDC

August 2022

<figure><img src="image_5.png" /></figure>

* Factors Associated with Delayed or Missed Second-Dose mRNA COVID-19 Vaccination among Persons >12 Years of Age, United States

* COVID-19 Vaccination Intent and Belief that Vaccination Will End the Pandemic

* Invasive Pneumococcal Disease and Long-Term Mortality Rates in Adults, Alberta, Canada

* Dog Ownership and Risk for Alveolar Echinococcosis, Germany

* Novel Chronic Anaplasmosis in Splenectomized Patient, Amazon Rainforest

* Transmissibility of SARS-CoV-2 B.1.1.214 and Alpha Variants during 4 COVID-19 Waves, Kyoto, Japan, January 2020–June 2021

* Culling of Urban Norway Rats and Carriage of *Bartonella* spp. Bacteria, Vancouver, British Columbia, Canada

* Zoonotic Threat of G4 Genotype Eurasian Avian-Like Swine Influenza A(H1N1) Viruses, China, 2020

* Increased Incidence of Invasive Pneumococcal Disease among Children after COVID-19 Pandemic, England

* Anthelmintic Baiting of Foxes against *Echinococcus multilocularis* in Small Public Area, Japan

* *Spiroplasma ixodetis* Infections in Immunocompetent and Immunosuppressed Patients after Tick Exposure, Sweden

* Child Melioidosis Deaths Caused by *Burkholderia pseudomallei*–Contaminated Borehole Water, Vietnam, 2019

* Association of Phylogenomic Relatedness among *Neisseria gonorrhoeae* Strains with Antimicrobial Resistance, Austria, 2016–2020

EMERGING
INFECTIOUS DISEASES

To revisit the August 2022 issue, go to:
https://wwwnc.cdc.gov/eid/articles/issue/28/8/table-of-contents