Climate Classification System–Based Determination of Temperate Climate Detection of Cryptococcus Gattii Sensu Lato

Climate Classification System–Based Determination of Temperate Climate Detection of Cryptococcus gattii sensu lato

Emily S. Acheson, Eleni Galanis, The Study Karen Bartlett, Brian Klinkenberg We used environmental isolations of C. gattii s.l. (i.e., de- tections in plant and soil samples) to map global distribu- We compared 2 climate classification systems describing geo- tion. Geographically defined human and animal records referenced environmental Cryptococcus gattii sensu lato isola- of C. gattii infection are not always accessible because of tions occurring during 1989–2016. Each system suggests the privacy restrictions. In addition, the dates and locations of fungus was isolated in temperate climates before the 1999 outbreak on Vancouver Island, British Columbia, Canada. How- C. gattii s.l. exposures are often uncertain because of the ever, the Köppen-Geiger system is more precise and should mobility of animals and humans and C. gattii’s long, unde- be used to define climates where pathogens are detected. termined incubation and latency periods (2). We extended the data of a database of globally georeferenced environmental isolations of C. gattii s.l. from the peer-reviewed global systematic framework is needed to define cli- literature (3) through November 2018 (Appendix Table). Amates where pathogens are detected. The 1999 cryp- We excluded studies in which only the country of sampling tococcal outbreak of the fungal species complex Crypto- was specified because many countries extend through mul- coccus gattii sensu lato on Vancouver Island (1), British tiple climates. We recorded the earliest year of isolation or, Columbia, Canada, was described as the first temperate cli- if a sampling year was not specified, the year of study pub- mate emergence of the pathogen because, before this event, lication. In total, we used 83 geographically unique coor- C. gattii s.l. was largely reported in areas described as tropi- dinates of C. gattii isolations occurring during 1989–2016. cal and subtropical (Appendix Figure, https://wwwnc.cdc. According to the solar definition (the predominant gov/EID/article/25/9/18-1884-App1.pdf). This assumption definition used in the C. gattii literature to describe iso- led to the belief that the Vancouver Island outbreak might lation climates), tropical, subtropical, and temperate re- be associated with a changing climate. The lack of precision gions are denoted by latitudinal boundaries (4). In con- and standardization of climate classification in the health trast, the Köppen-Geiger system (5) uses precipitation, literature makes comparing emergence areas around the temperature, and vegetation traits to produce 5 main cli- world and determining why or how the organism emerged mate groups (tropical or equatorial, arid, temperate, con- in that area difficult. tinental, and polar) and subgroups and is the most widely The lack of consensus might largely be rooted in the used climate classification system by researchers, includ- lack of a global systematic framework to define climates ing medical geographers, worldwide (6). For the solar where pathogens are detected. Specifically, a standardized definition of climate, we set the latitudinal boundaries of definition of tropical, subtropical, and temperate to com- the tropics at 23.4 degrees north and south of the equator, pare pathogen isolation areas worldwide is unavailable. the area of the subtropics as the tropical extent to 35 or 40 Here, we compare the 2 solar climate definitions and the degrees north and south of the equator, and the temperate Köppen-Geiger climate classification system to determine area as the subtropical extent to 66.5 degrees north and whether the 1999 Vancouver Island outbreak was the first- south of the equator (4,7). For the Köppen-Geiger system, ever detection of C. gattii s.l. in a temperate environment we used a map depicting the climate characteristics ob- and which system should be used for a global systematic served during 1976–2000 with a spatial resolution of 0.5 climate classification framework. degrees (http://koeppen-geiger.vu-wien.ac.at/shifts.htm). Using ArcMap version 10.5.1 (ESRI 2017, https://www. Author affiliations: University of British Columbia, Vancouver, esri.com), we overlaid the isolation coordinates of C. gat- British Columbia, Canada (E.S. Acheson, E. Galanis, K. Bartlett, tii on each map and extracted the corresponding climate B. Klinkenberg); British Columbia Centre for Disease Control, classifications. If exact coordinates of the sample were not Vancouver (E. Galanis) specified, we used ArcMap to estimate coordinates for the DOI: https://doi.org/10.3201/eid2509.181884 centroid of the park, city, or town where the sampling

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 9, September 2019 1723 DISPATCHES occurred. We first compared the climates assigned to Geiger system, the Vancouver Island outbreak areas have isolations by classification system and then determined 2 different types of temperate climates: temperate oceanic which positive sampling years had >1 isolation in a tem- and warm summer Mediterranean, that is, precipitation perate region and how many of these years preceded 1999. conditions that range from dry summers to fully humid The solar definition classified the 83 environmental year-round with warm summer temperatures (5). Accord- C. gattii s.l. isolations as tropical, subtropical, or temper- ing to the more restrictive solar definition of temperate, the ate (Figure 1). By comparison, the Köppen-Geiger system environmental C. gattii s.l. isolation coordinates from only classified these same isolations into 11 different climate 1 year before 1999 could be classified as temperate (Fig- subgroups (Figure 1). Both systems identified >1 temper- ure 2). By contrast, the Köppen-Geiger system classified ate-climate environmental C. gattii isolation (8) before the coordinates of environmental isolations from 7 differ- 1999 (Figure 2). ent years before 1999 as temperate (Figure 2; Appendix Both variations of the solar definition and the Köppen- Table). These isolation coordinates included areas in Cali- Geiger system classified the environmental samples of C. fornia (9,10) and southwestern and southeastern Australia gattii s.l. isolated on Vancouver Island during the outbreak (9,11). The solar definition of climate largely categorized as temperate (Appendix Table). According to the Köppen- these areas as tropical or subtropical (Appendix Table), but

Figure 1. Global environmental isolations of Cryptococcus gattii sensu lato, 1989–2016. We mapped 83 unique geographic coordinates of C. gattii s.l. isolations and labeled them according to their Köppen-Geiger climate classification. Overlapping symbols of the same Köppen-Geiger climate classification (where isolations were 0–200 km apart) were removed for easier visualization.The solar definition of the tropics is shown as the semitransparent red area extending from the equator to 23.4 degrees north and south of the equator, the subtropics as the yellow area extending from the tropics to either 35 (solid line) or 40 (dashed line) degrees north and south of the equator, and the temperate zone as the green area extending from the subtropics to 66.5 degrees north and south of the equator.

1724 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 9, September 2019 Temperate Climate Detection of C. gattii s.l.

Figure 2. Timeline of environmental Cryptococcus gattii sensu lato isolations in temperate climates, by climate definition, 1989–2016. Gray squares indicate years in which >1 isolate from a temperate climate was obtained, and white squares indicate years in which no such isolate was obtained. the Köppen-Geiger system labeled them as temperate oce- exact coordinates of C. gattii sampling, and others pro- anic and warm summer Mediterranean. vided a park or city name. Providing the exact coordinates offers the greatest certainty of a detection location and bet- Conclusions ter precision in climate classification. Both versions of the solar definition and the Köppen- By using C. gattii s.l. as an example for mapping Geiger system suggest that the Vancouver Island out- georeferenced pathogen isolations worldwide, we demon- break was not the first-ever temperate detection ofC. gat- strated the opportunity to improve pathogen monitoring tii s.l. However, in terms of geographic scale, the solar through the development of a standardized global climate definition of tropical, subtropical, and temperate are too classification framework. Using more spatially specific cli- coarse for the purposes of classifying or describing areas mate classification methods, such as the Köppen-Geiger of local clinical, veterinary, or environmental isolations system used by medical geographers, coupled with the of C. gattii s.l. or any other environmentally contracted continued reporting of pathogen isolation locations, will pathogen. Species, including pathogenic species, can live improve comparability of pathogen detection in new natu- within geographically smaller refugia that maintain their ral environments. climatic and biological needs across larger landscapes and solar boundaries, depending on topography, micro- Acknowledgments climates, and habitat fragmentation (12,13). Although the The authors thank Richard Malik, Mark Krockenberger, Peter Köppen-Geiger system still generalizes across precipita- Irwin, and Cristy Secombe for their expert input regarding tion, temperature, and vegetation, the system accounts for C. gattii outbreaks and emergence in the environment in Australia. more environmental variation and provides temperature We also thank Mike Jerowsky, Peter Whitman, and José Aparicio and precipitation limits for each climate subtype. The for their help with the figures and edits to the manuscript. Köppen-Geiger system is the most widely used climate Financial support for this project was provided by a Vanier classification system worldwide5 ( ) and also provides Canada Graduate Scholarship for the Natural Sciences and projected maps for future climate shifts, making this sys- Engineering Research Council of Canada, a University of British tem ideal as a global systematic framework for tracking Columbia Four-Year Fellowship, and a Killam Doctoral the climates of pathogen detection. We, therefore, pro- Scholarship to E.S.A. pose the use of the Köppen-Geiger system, as opposed to either of the overgeneralized solar definitions, for the sake of precision and consistency across global records when About the Author characterizing pathogen detection areas. Ms. Acheson is a doctoral candidate in the Geography One limitation of our study was dependence on the Department at the University of British Columbia, Vancouver, reporting of environmental C. gattii samples in the English British Columbia, Canada. Her primary research interests are in language peer-reviewed literature. As a result, our find- infectious disease emergence and disease vector ecology. ings are an underrepresentation of the full global extent of C. gattii s.l. in the environment. Other evidence exists for the emergence of C. gattii s.l. in temperate climates References 1. Stephen C, Lester S, Black W, Fyfe M, Raverty S. Multispecies before 1999. For example, in addition to the environmen- outbreak of cryptococcosis on southern Vancouver Island, British tal isolations made in Busselton, Western Australia, Aus- Columbia. Can Vet J. 2002;43:792–4. tralia (9), in 1993, multiple C. gattii infections in animals 2. Hagen F, Colom MF, Swinne D, Tintelnot K, Iatta R, Montagna were reported in southwestern Australia, including Perth, MT, et al. Autochthonous and dormant Cryptococcus gattii infections in Europe. Emerg Infect Dis. 2012;18:1618–24. before 1999 (14). Both Busselton and Perth fall within a https://doi.org/10.3201/eid1810.120068 temperate Köppen-Geiger climate (5) (Figure 1). Another 3. Acheson ES, Galanis E, Bartlett K, Mak S, Klinkenberg B. limitation was variability in the descriptions of pathogen Searching for clues for eighteen years: deciphering the ecological detection areas. For example, some studies provided the determinants of Cryptococcus gattii on Vancouver Island, British

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 9, September 2019 1725 DISPATCHES

Columbia. Med Mycol. 2018;56:129–44. https://doi.org/10.1093/ 11. Pfeiffer TJ, Ellis DH. Environmental isolation ofCryptococcus mmy/myx037 neoformans var. gattii from Eucalyptus tereticornis. J Med 4. Corlett RT. Where are the subtropics? Biotropica. 2013;45:273–5. Vet Mycol. 1992;30:407–8. https://doi.org/10.1080/ https://doi.org/10.1111/btp.12028 02681219280000541 5. Peel MC, Finlayson BL, McMahon TA. Updated world map of 12. Varner J, Dearing MD. The importance of biologically relevant the Koppen-Geiger climate classification. Hydrol Earth Syst Sci. microclimates in habitat suitability assessments. 2007;11:1633–44. https://doi.org/10.5194/hess-11-1633-2007 PLoS One. 2014;9:e104648. https://doi.org/10.1371/journal. 6. Polack S, Brooker S, Kuper H, Mariotti S, Mabey D, Foster A. pone.0104648 Mapping the global distribution of trachoma. Bull World Health 13. Jackson MM, Gergel SE, Martin K. Effects of climate change on Organ. 2005;83:913–9. habitat availability and configuration for an endemic coastal alpine 7. Monson RK. Ecology of temperate forests. Ecology and the bird [Erratum: PLoS ONE. 2015;11:e0146838]. environment. New York: Springer New York; 2014. p. 273–96. PLoS One. 2015;10:e0142110. https://doi.org/10.1371/journal. 8. Montagna MT, Viviani MA, Pulito A, Aralla C, Tortorano AM, pone.0142110 Fiore L, et al. Cryptococcus neoformans var. gattii in Italy. Note II. 14. McGill S, Malik R, Saul N, Beetson S, Secombe C, Environment investigation related to an autochthonous clinical case Robertson I, et al. Cryptococcosis in domestic animals in in Apulia. J Mycol Med. 1997;7:93–6. Western Australia: a retrospective study from 1995–2006. 9. Sorrell TC, Chen SCA, Ruma P, Meyer W, Pfeiffer TJ, Ellis DH, Med Mycol. 2009;47:625–39. https://doi. et al. Concordance of clinical and environmental isolates of org/10.1080/13693780802512519 Cryptococcus neoformans var. gattii by random amplification of polymorphic DNA analysis and PCR fingerprinting. J Clin Address for correspondence: Emily S. Acheson, University of Microbiol. 1996;34:1253–60. British Columbia, Lab for Advanced Spatial Analysis, Department of 10. Pfeiffer T, Ellis D. Environmental isolation ofCryptococcus neoformans gattii from California. J Infect Dis. 1991;163:929–30. Geography, 1984 West Mall, Rm 210J, Vancouver, British Columbia https://doi.org/10.1093/infdis/163.4.929 V6T 1Z2, Canada; email: [email protected]

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1726 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 9, September 2019

Article DOI: https://doi.org/10.3201/eid2509.181884 Climate Classification System–Based Determination of Temperate Climate Detection of Cryptococcus gattii sensu lato

Appendix

Methods for Updated Literature Review

To assess how historical (i.e., pre-1999) areas of C. gattii endemicity have been described in the literature, we searched for peer-reviewed English articles published from January 1970 through November 2018 that directly analyzed or reviewed the ecology and/or geographic distribution of C. gattii. We searched Google Scholar (http://scholar.google.com), ScienceDirect (http://www.sciencedirect.com), and Web of Science (http://thomsonreuters.com/thomsonreuters-web-of-science). For species, we selected the inputs “either/or” for “Cryptococcus gattii,” “Cryptococcus neoformans var. gattii,” or “Cryptococcus bacillisporus” in the search engine, where the last was considered synonymous with C. gattii in the late 1970s and early 1980s (1); for ecologic or geographic studies, we selected “either/or” for “ecolog*,” “geograph*,” “distribution,” and “niche.” In addition to assessing how historical areas of endemicity of C. gattii were described, we also identified which studies were used as the original sources of these statements.

We originally found 105 articles that studied C. gattii ecology or geographic distributions, but further narrowed our results down to 73 articles that directly discussed areas of C. gattii endemicity before the Vancouver Island outbreak. Articles were classed as “restricted” if they worded C. gattii distributions as “restricted to,” “only found in,” or “confined to” (sub)tropical areas, or variations thereof. Articles were classed as “general” if they worded C. gattii distributions as “mainly,” “predominantly,” or “primarily” in (sub)tropical areas, or variations thereof.

Of the 73 articles reviewed, 35 studies contained statements claiming C. gattii was restricted to the (sub)tropics before the Vancouver Island outbreak (i.e., “restricted” statements),

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while 38 studies also acknowledged other historical areas of endemicity outside the (sub)tropics (i.e., “general” statements) (Appendix Figure). The same studies were often cited as the sources of both statements. For example, 2 seminal papers published in 1984 on global Cryptococcus neoformans and C. gattii distributions (2,3) were primarily or secondarily cited in 24 of the 35 studies making “restricted” statements but were also cited in 10 of the 38 studies making “general” statements. The authors demonstrated a higher prevalence of C. gattii in (sub)tropical areas compared to temperate ones but stated the fungus was not entirely restricted to these warmer areas.

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46. Hagen F, Chowdhary A, Prakash A, Yntema J-B, Meis JF. Molecular characterization of Cryptococcus gattii genotype AFLP6/VGII isolated from woody debris of divi-divi (Caesalpinia coriaria), Bonaire, Dutch Caribbean. Rev Iberoam Micol. 2014;31:193–6. PubMed https://doi.org/10.1016/j.riam.2013.10.007

47. Cogliati M, D’Amicis R, Zani A, Montagna MT, Caggiano G, De Giglio O, et al. Environmental distribution of Cryptococcus neoformans and C. gattii around the Mediterranean basin. FEMS Yeast Res. 2016;16:fow045. PubMed https://doi.org/10.1093/femsyr/fow045

48. Linares C, Colom MF, Torreblanca M, Esteban V, Romera Á, Hagen F. Environmental sampling of Ceratonia siliqua (carob) trees in Spain reveals the presence of the rare Cryptococcus gattii genotype AFLP7/VGIV. Rev Iberoam Micol. 2015;32:269–72. PubMed https://doi.org/10.1016/j.riam.2014.11.002

49. Brito-Santos F, Barbosa GG, Trilles L, Nishikawa MM, Wanke B, Meyer W, et al. Environmental isolation of Cryptococcus gattii VGII from indoor dust from typical wooden houses in the deep Amazonas of the Rio Negro basin. PLoS One. 2015;10:e0115866. PubMed https://doi.org/10.1371/journal.pone.0115866

50. Khayhan K, Hagen F, Norkaew T, Puengchan T, Boekhout T, Sriburee P. Isolation of Cryptococcus gattii from a Castanopsis argyrophylla tree hollow (Mai-Kaw), Chiang Mai, Thailand. Mycopathologia. 2017;182:365–70. PubMed https://doi.org/10.1007/s11046-016-0067-7

51. Hagen F, Khayhan K, Theelen B, Kolecka A, Polacheck I, Sionov E, et al. Recognition of seven species in the Cryptococcus gattii/Cryptococcus neoformans species complex. Fungal Genet Biol. 2015;78:16–48. PubMed https://doi.org/10.1016/j.fgb.2015.02.009

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Appendix Table. Unique georeferenced global environmental isolations of Cryptococcus gattii (n = 83) listed in chronologic order, 1989–2016* With With Sample Earliest Mating ±40° ±35° KG KG Country Area year(s) Longitude Latitude year Genotype type Species limits limits label limits Ref. Australia Balranald, NSW 1989 143.5633 34.6394 1989 – – – ST ST BSk AR (4) Australia Balranald, NSW 1989–1990 143.5633 34.6394 1989 AFLP4B/VGIb – C. gattii s.s. ST ST BSk AR (5) Australia Balranald, NSW 1989–1996 143.5633 34.6394 1989 AFLP4/VGI α, a C. gattii s.s. ST ST BSk AR (6) Australia Hay, NSW 1989 144.5803 34.1573 1989 – – – ST ST BSk AR (4) Australia Hay, NSW 1989–1990 144.5803 34.1573 1989 AFLP4/VGI α C. gattii s.s. ST ST BSk AR (6) Australia Barossa Valley (Barossa Reservoir 1989 138.95 34.5333 1989 – – – ST ST Csa TM (4) and Nuriootpa), SA Australia Barossa Valley, SA 1989–1992 138.95 34.5333 1989 AFLP4A/VGIa – C. gattii s.s. ST ST Csa TM (5) United Fort Point, San Francisco, 1990 122.477 37.81022 1990 AFLP6A/VGIIa – C. deuterogattii ST TM Csb TM (7) States California Australia Tocumwal, NSW 1991 145.569 35.8122 1991 AFLP4/VGI – C. gattii s.s. ST TM BSk AR (5) Australia Currumbin, Gold Coast, QLD 1991 153.4667 28.1333 1991 – – – ST ST Cfa TM (8) Australia Currumbin, Gold Coast, QLD 1991–1993 153.4667 28.1333 1991 AFLP4/VGI – C. gattii s.s. ST ST Cfa TM (5) Australia Mt. Annan, Greater Sydney, NSW 1991 150.7598 34.0529 1991 – – – ST ST Cfb TM (8) Australia Mt. Annan, Greater Sydney, NSW 1991–1994 150.7598 34.0529 1991 AFLP4/VGI – C. gattii s.s. ST ST Cfb TM (5) Australia Busselton, WA 1993 115.3708 33.6848 1993 AFLP6/VGII – C. deuterogattii ST ST Csb TM (5) Brazil Rio de Janeiro <1993 43.16667 22.9 1993 AFLP4/VGI – C. gattii s.s. T T Am T (9) Brazil Teresina 1993 42.8485 5.30175 1993 AFLP6/VGII α C. deuterogattii T T Aw T (10) Brazil Teresina 1993–1997 42.8485 5.30175 1993 AFLP6/VGII α C. deuterogattii T T Aw T (11) India Ferozepur 1995–1996 74.56 30.89 1995 – – – ST ST BSh AR (12) Australia Gold Coast, QLD 1996 153.4309 28.0003 1996 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (6) Australia Adelaide, SA 1996 138.5986 34.9287 1996 AFLP4/VGI α C. gattii s.s. ST ST Csa TM (6) Brazil Ibirapuera Park, Sao Paulo 1996–1997 46.6581 23.6022 1996 – – – ST ST Cfa TM (13) Italy Apulia 1996 16.619 40.985 1996 – – – TM TM Cfa TM (14) United San Diego Zoo area, San Diego, <1996 117.157 32.71533 1996 AFLP4/VGI – C. gattii s.s. ST ST BSk AR (5) States California AFLP5/VGIII C. bacillisporus Australia Coffs Harbour, NSW 1997–2000 153.10 30.30 1997 – α – ST ST Cfa TM (15) Australia Port Macquarie, NSW 1997–2000 152.92 31.44 1997 – α – ST ST Cfa TM (15) Australia St. Ives, Sydney, NSW 1997–1998 151.1667 33.7333 1997 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (6) Australia Eastern Sydney, NSW 1997–2000 151.20 33.86 1997 – α, a – ST ST Cfa TM (15) Colombia Cúcuta 1997 72.5 7.9 1997 – – – T T Am T (16) Australia Renmark, SA 1998 140.747 34.177 1998 AFLP4/VGI α, a C. gattii s.s. ST ST BSk AR (6) Australia Breeza, NSW 1998 150.4667 31.25 1998 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (6) Australia Pilliga, NSW 1998 148.9 30.35 1998 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (6) Australia Port Macquarie, NSW 1998 152.9167 31.4167 1998 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (6) Brazil Ilha de Maracá 1998–1999 61.6667 3.41667 1998 AFLP6/VGII a C. deuterogattii T T Am T (17) Egypt Qutur and Tanta areas, Gharbia 1998 30.95614 30.97225 1998 – – – ST ST BWh AR (18) Governorate India Delhi (northwestern India) 1999–2000 77.22897 28.65381 1999 – – – ST ST BSh AR (19) Mexico Mexico City <1999 99.1277 19.42847 1999 – – – T T Cwb TM (20) Australia Glenbrook, Blue Mountains 2000 150.4731 33.6562 2000 AFLP4/VGI α C. gattii s.s. ST ST Cfb TM (21) National Park, Sydney Argentina Buenos Aires City 2001 58.3772 34.6131 2001 – – – ST ST Cfa TM (22)

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With With Sample Earliest Mating ±40° ±35° KG KG Country Area year(s) Longitude Latitude year Genotype type Species limits limits label limits Ref. Argentina Rep. de Chile Park, de los Patricios 2002 58.3772 34.6131 2002 AFLP4/VGI – C. gattii s.s. ST ST Cfa TM (23) Park, Centenario Park, and N. Avellaneda Park in Buenos Aires City Canada Cameron Lake, B.C. 2002 124.619 49.29133 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Canada Courtenay, B.C. 2002 124.994 49.68657 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Canada Duncan, B.C. 2002 123.703 48.78293 2002 AFLP4/VGI – C. gattii s.s. TM TM Csb TM (24) AFLP6B/VGIIb C. deuterogattii Canada MacMillan Park, Cathedral Grove, 2002 124.669 49.28293 2002 AFLP6A/VGIIa α C. deuterogattii TM TM Csb TM (25) B.C. Canada Nanaimo, B.C. 2002 123.936 49.16634 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Canada Parksville, B.C. 2002 124.314 49.32 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Canada Port Alberni, B.C. 2002 124.808 49.234 2002 AFLP6A/VGIIa, – C. deuterogattii TM TM Csb TM (24) AFLP6B/VGIIb Canada Rathtrevor Beach Provincial Park, 2002 124.266 49.32096 2002 AFLP6A/VGIIa, α C. deuterogattii TM TM Csb TM (25) Parksville, B.C. AFLP6B/VGIIb Canada Victoria, B.C. 2002 123.369 48.43294 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Colombia Cali 2002–2003 75.68333 5.0 2002 AFLP6/VGII a C. deuterogattii T T Af T (26) Colombia Cúcuta 2002–2003 72.01667 6.933333 2002 AFLP4/VGI α C. gattii s.s. T T Cfb TM (26) AFLP5/VGIII C. bacillisporus VGIV (AFLP type – not specified) Colombia Bogota 2002–2003 74.08333 4.6 2002 AFLP6/VGII a C. deuterogattii T T Cfb TM (26) Colombia Cundinamarca 2002–2003 73.05 3.666667 2002 AFLP6/VGII a C. deuterogattii T T Am T (26) VGIV (AFLP type α – not specified) Colombia Medellin 2002–2003 73.88333 5.433333 2002 AFLP6/VGII a C. deuterogattii T T Cfb TM (26) India Jabalpur City (Central India) 2002–2004 79.95006 23.16697 2002 – – – T T Csa TM (27) United Lynden, Washington 2002–2006 122.452 48.9465 2002 AFLP6A/VGIIa – C. deuterogattii TM TM Cfb TM (24) States Australia Mt. Druitt, Sydney, NSW <2003 150.8167 33.7667 2003 AFLP4/VGI α C. gattii s.s. ST ST Cfa TM (28) AFLP6/VGII α C. deuterogattii Colombia La Calera 2003 73.9693 4.72069 2003 – a – T T Cfb TM (29) Canada Saltspring Island, B.C. 2004–2005 123.408 48.769 2004 AFLP4/VGI – C. gattii s.s. TM TM Csb TM (30) Canada Hornby Island 2005 124.667 49.525 2005 AFLP6A/VGIIa – C. deuterogattii TM TM Csb TM (24) Argentina Resistencia, Chaco 2006 58.9839 27.4606 2006 AFLP5/VGIII – C. bacillisporus ST ST Cfa TM (31) Canada Elk Falls Provincial Park, Campbell 2006 125.251 50.026 2006 – – – TM TM Cfb TM (24) River Argentina Parque España, La Paz, Entre Rios 2007 59.6434 30.7437 2007 AFLP4/VGI – C. gattii s.s. ST ST Cfa TM (31) Argentina Rosario, Santa Fe 2007 60.6393 32.9468 2007 AFLP4/VGI – C. gattii s.s. ST ST Cfa TM (31) Brazil Botafogo district, Rio de Janeiro 2008–2010 43.1856 22.969 2008 AFLP4/VGI – C. gattii s.s. T T Am T (32) Colombia Cúcuta 2008–2009 72.51 7.913333 2008 AFLP4/VGI a C. gattii s.s. T T Cfb TM (33) AFLP5/VGIII α C. bacillisporus Brazil Belem, Para <2009 48.50417 1.45556 2009 AFLP6/VGII – C. deuterogattii T T Am T (34) India Guindy National Park, Chennai, <2009 80.21941 12.99668 2009 – – – T T Aw T (35) South India

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With With Sample Earliest Mating ±40° ±35° KG KG Country Area year(s) Longitude Latitude year Genotype type Species limits limits label limits Ref. Italy Reggio Calabria (southern Italy) 2009 15.65 38.07 2009 AFLP4/VGI α C. gattii s.s. ST TM Csa TM (36) Puerto Rico Guanica Dry Forest <2010 66.908 17.97163 2010 AFLP6/VGII α C. deuterogattii T T Aw T (37) AFLP7/VGIV α C. tetragattii United Near Silver Falls State Park, 2010–2011 122.6336 44.90544 2010 AFLP4/VGI, – C. gattii s.s., C. TM TM Csb TM (38) States Oregon (based on Figure 1 in main AFLP6A/VGIIa, deuterogattii text) AFLP6B/VGIIb, AFLP6C/VGIIc The Berg en Dal 2011 5.916667 51.82167 2011 AFLP4/VGI α C. gattii s.s. TM TM Cfb TM (39) Netherlands Tunisia Sfax region (southern Tunisia) <2011 10.76028 34.74056 2011 – α – ST ST BSh AR (40) United Los Angeles, California 2011–2012 118.244 34.05223 2011 AFLP4/VGI α C. gattii s.s. ST ST Csb TM (41) States AFLP5A/VGIIIa α AFLP5B/VGIIIb a, α Botswana Francistown 2012 27.50788 21.17 2012 – a, α – T T BSh AR (42) Gaborone 2012 25.933 24.651 2012 – α – ST ST BSh AR Maun 2012 23.415 20.007 2012 – a, α – T T BSh AR Brazil Manaus 2012–2014 60.052 3.109 2012 AFLP6/VGII α C. deuterogattii T T Af T (43) Kenya Nairobi 2012–2013 36.83333 1.28333 2012 AFLP4/VGI – C. gattii s.s. T T Cfb TM (44) Spain Alicante <2012 0.48149 38.34517 2012 AFLP4/VGI α C. gattii s.s. ST TM BSk AR (45) Spain Barcelona <2012 2.158987 41.38879 2012 AFLP4/VGI α C. gattii s.s. TM TM Csa TM (45) Bonaire Lagun Goto, Rincon village, Hato 2013 68.3258 12.21501 2013 AFLP6/VGII α C. deuterogattii T T BSh† AR (46) village Greece Athens 2013 23.71622 37.97945 2013 AFLP4/VGI α C. gattii s.s. ST TM Csa TM (47) Greece Salamina Island 2013 23.50677 37.92189 2013 AFLP4/VGI α C. gattii s.s. ST TM Csa TM (47) Italy Ragalna, Catania 2013 15.08233 37.43505 2013 AFLP4/VGI α C. gattii s.s. ST TM Csa TM (47) Italy Route Brindisi-Fasano 2013 17.69123 40.60921 2013 AFLP4/VGI a C. gattii s.s. TM TM Csa TM (47) Italy Route Gallipoli-Collepasso 2013 17.99538 40.0559 2013 AFLP4/VGI a C. gattii s.s. TM TM Csa TM (47) Spain El Perello, Tarragona 2013 0.71335 40.87642 2013 AFLP7/VGIV α C. tetragattii TM TM Csa TM (47, 48) Spain Campello, Alicante 2014 0.398 38.429 2014 AFLP4/VGI α C. gattii s.s. ST TM BSk AR (47) Spain Mendivil, Navarra 2014 1.451 42.66366 2014 AFLP4/VGI α C. gattii s.s. TM TM Cfb TM (47) Brazil Santa Isabel do Rio Negro, <2015 65.01889 0.41417 2015 AFLP6/VGII α, a C. deuterogattii T T Af T (49) Amazonas state Thailand Queen Sirikit Botanic Garden, <2016 98.86 18.89953 2016 AFLP4/VGI – C. gattii s.s. T T Aw T (50) Chiang Mai Province *For longitude, positive numbers refer to degrees east of the prime meridian and negative numbers refer to degrees west of the prime meridian. For latitude, positive numbers refer to degrees north of the equator and negative numbers refer to degrees south of the equator. The latitudinal boundaries of ±40° and ±35° refer to degrees north (+) and south (-) of the equator. For each isolation, we recorded the country and area of isolation, the range of years when the isolations were collected (if the earliest year yielding positive isolations was provided, this would be the start of the range), the coordinates where the isolation was found, the earliest possible year of collection recorded for this analysis, and the climate classifications for each isolation, as designated by solar definitions with ±40° and ±35° limits as well as by the KG system. Genotype (including subtype), mating type, and species (51) are also provided, if specified by the study. It is important to note that some studies have only used phenotypic methods (e.g., CGB agar) to differentiate C. gattii sensu lato from C. neoformans s.l.. This is, however, not always a reliable method. Thus, data from this list need to be interpreted with caution. AR, arid; B.C., British Columbia; KG, Köppen-Geiger; NSW, New South Wales; QLD, Queensland; ref., reference; SA, South Australia; ST, subtropical; T, tropical; TM, temperate; WA, Western Australia. †The KG system does not classify the island of Bonaire (Dutch Caribbean) in its map of observed climates during 1976–2000. We, therefore, obtained the classification of Bonaire from (52) in addition to our own categorization using the KG climate classification criteria (53) and present-day climate averages of the island (https://www.ncdc.noaa.gov/).

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Appendix Figure. Publications from 1984 through November 2018 discussing pre-1999 areas of endemicity of Cryptococcus gattii (n = 73). Publications containing statements claiming C. gattii was strictly found in tropical and subtropical areas before the 1999 Vancouver Island outbreak were labeled as “restricted” (n = 35), while those containing statements that also acknowledged other areas of endemicity before the 1999 Vancouver Island outbreak were labeled as “general” (n = 38).

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