Cryptococcus gattii VGIII Isolates Causing Infections in HIV/AIDS Patients in Southern California: Identification of the Local Environmental Source as Arboreal

PLoS Pathogens

Volumn 10, Issue 8, 2014, Pages

  Springer, Deborah J ^a   Billmyre, R Blake ^a   Filler, Elan E ^b   Voelz, Kerstin ^c,d   Pursall, Rhiannon ^c   Mieczkowski, Piotr A ^e   Larsen, Robert A ^f   Dietrich, Fred S ^a   May, Robin C ^c,d   Filler, Scott G ^b   Heitman, Joseph ^a  

^a DUKE UNIVERSITY MEDICAL CENTER   (United States)

^b UNIVERSITY OF CALIFORNIA   (United States)

^c UNIVERSITY OF BIRMINGHAM   (United Kingdom)

^d QUEEN ELIZABETH HOSPITAL   (United Kingdom)

^e UNIVERSITY OF NORTH CAROLINA SCHOOL OF MEDICINE   (United States)

^f UNIVERSITY OF SOUTHERN CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

AMPHOTERICIN B; FLUCONAZOLE; FLUCYTOSINE; KETOCONAZOLE;

ACQUIRED IMMUNE DEFICIENCY SYNDROME; ANIMAL EXPERIMENT; ANIMAL MODEL; ANTIFUNGAL SUSCEPTIBILITY; ARBOREAL SPECIES; ARTICLE; BACTERIUM ISOLATE; CELL PROLIFERATION ASSAY; CRYPTOCOCCUS GATTII; DRUG SENSITIVITY; ENVIRONMENTAL DISEASE; FLUORESCENCE ACTIVATED CELL SORTING; GENE LOCUS; GENE SEQUENCE; GENETIC ANALYSIS; GENETIC DIFFERENCE; HAPLOTYPE; HUMAN; HUMAN IMMUNODEFICIENCY VIRUS INFECTED PATIENT; INTROGRESSION; MOUSE; MULTILOCUS SEQUENCE TYPING; NONHUMAN; PATHOGENESIS; PINE; PLOIDY; UNITED STATES; ANIMAL; BAGG ALBINO MOUSE; CALIFORNIA; CELL SEPARATION; COMPLICATION; CRYPTOCOCCOSIS; DISEASE MODEL; FEMALE; GENETICS; HUMAN IMMUNODEFICIENCY VIRUS INFECTION; MALE; MICROBIOLOGY; PHYLOGENY; POLYMERASE CHAIN REACTION; TREE;

ACQUIRED IMMUNODEFICIENCY SYNDROME; ANIMALS; CALIFORNIA; CELL SEPARATION; CRYPTOCOCCOSIS; CRYPTOCOCCUS GATTII; DISEASE MODELS, ANIMAL; FEMALE; HIV INFECTIONS; HUMANS; MALE; MICE; MICE, INBRED BALB C; PHYLOGENY; POLYMERASE CHAIN REACTION; SOIL MICROBIOLOGY; TREES;

EID: 84912121107     PISSN: 15537366     EISSN: 15537374     Source Type: Journal    
DOI: 10.1371/journal.ppat.1004285     Document Type: Article

References (97)

1. Reed C, Biggerstaff M, Finelli L, Koonin LM, Beauvais D, et al. (2013) Novel framework for assessing epidemiologic effects of influenza epidemics and pandemics. Emerg Infect Dis 19: 85–91.
2. Dawood FS, Iuliano AD, Reed C, Meltzer MI, Shay DK, et al. (2012) Estimated global mortality associated with the first 12 months of 2009 pandemic influenza A H1N1 virus circulation: a modelling study. Lancet Infect Dis 12: 687–695.
3. Jutla A, Whitcombe E, Hasan N, Haley B, Akanda A, et al. (2013) Environmental factors influencing epidemic cholera. Am J Trop Med Hyg 89: 597–607.
4. Christian KA, Ijaz K, Dowell SF, Chow CC, Chitale RA, et al. (2013) What we are watching–five top global infectious disease threats, 2012: a perspective from CDC's Global Disease Detection Operations Center. Emerg Health Threats J 6: 20632.
5. de Hoog GS, Guarro J, Gene J, Figueras MJ (2000) Atlas of Clinical Fungi. Utrecht: Centraalbureau voor Schimmelcultures/Universitat Rovira i Virgili.
6. Fisher FS, Bultman MW, Johnson SM, Pappagianis D, Zaborsky E, (2007) Coccidioides niches and habitat parameters in the southwestern United States: a matter of scale. Ann N Y Acad Sci 1111: 47–72.
7. Pappagianis D, (1988) Epidemiology of coccidioidomycosis. Curr Top Med Mycol 2: 199–238.
8. Howard DH, (1984) The epidemiology and ecology of blastomycosis, coccidioidomycosis and histoplasmosis. Zentralbl Bakteriol Mikrobiol Hyg A 257: 219–227.
9. Barker BM, Jewell KA, Kroken S, Orbach MJ, (2007) The population biology of coccidioides: epidemiologic implications for disease outbreaks. Ann N Y Acad Sci 1111: 147–163.
10. Neblett Fanfair R, Benedict K, Bos J, Bennett SD, Lo YC, et al. (2012) Necrotizing cutaneous mucormycosis after a tornado in Joplin, Missouri, in 2011. N Engl J Med 367: 2214–2225.
11. Chamany S, Mirza SA, Fleming JW, Howell JF, Lenhart SW, et al. (2004) A large histoplasmosis outbreak among high school students in Indiana, 2001. Pediatr Infect Dis J 23: 909–914.
12. Cairns L, Blythe D, Kao A, Pappagianis D, Kaufman L, et al. (2000) Outbreak of coccidioidomycosis in Washington state residents returning from Mexico. Clin Infect Dis 30: 61–64.
13. Brown GD, Denning DW, Gow NA, Levitz SM, Netea MG, et al. (2012) Hidden killers: human fungal infections. Sci Transl Med 4: 165rv113.
14. Hagen F, Colom MF, Swinne D, Tintelnot K, Iatta R, et al. (2012) Autochthonous and dormant Cryptococcus gattii infections in Europe. Emerging Infectious Diseases 18: 1618–1624.
15. Meyer W, Aanensen DM, Boekhout T, Cogliati M, Diaz MR, et al. (2009) Consensus multi-locus sequence typing scheme for Cryptococcus neoformans and Cryptococcus gattii. Med Mycol 47: 1–14.
16. Cogliati M, (2013) Global molecular epidemiology of Cryptococcus neoformans and Cryptococcus gattii: An atlas of the molecular types. Scientifica 2013: 23.
17. Park BJ, Wannemuehler KA, Marston BJ, Govender N, Pappas PG, et al. (2009) Estimation of the current global burden of cryptococcal meningitis among persons living with HIV/AIDS. AIDS 23: 525–530.
18. Pyrgos V, Seitz AE, Steiner CA, Prevots DR, Williamson PR, (2013) Epidemiology of cryptococcal meningitis in the US: 1997–2009. PLOS ONE 8: e56269.
19. Dixon DM, McNeil MM, Cohen ML, Gellin BG, La Montagne JR, (1996) Fungal infections: a growing threat. Public Health Rep 111: 226–235.
20. Springer DJ, Chaturvedi V, (2010) Projecting global occurrence of Cryptococcus gattii. Emerg Infect Dis 16: 14–20.
21. Byrnes EJ, 3rdLi W, Lewit Y, Ma H, Voelz K, et al. (2010) Emergence and pathogenicity of highly virulent Cryptococcus gattii genotypes in the Northwest United States. PLOS Pathogens 6: e1000850.
22. Byrnes EJ, 3rdLi W, Ren P, Lewit Y, Voelz K, et al. (2011) A diverse population of Cryptococcus gattii molecular type VGIII in Southern Californian HIV/AIDS patients. PLOS Pathogens 7: e1002205.
23. Meyer W, Castaneda A, Jackson S, Huynh M, Castaneda E, (2003) Molecular typing of IberoAmerican Cryptococcus neoformans isolates. Emerging Infectious Diseases 9: 189–195.
24. Ngamskulrungroj P, Gilgado F, Faganello J, Litvintseva AP, Leal AL, et al. (2009) Genetic diversity of the Cryptococcus species complex suggests that Cryptococcus gattii deserves to have varieties. PLOS ONE 4: e5862.
25. Fraser JA, Giles SS, Wenink EC, Geunes-Boyer SG, Wright JR, et al. (2005) Same-sex mating and the origin of the Vancouver Island Cryptococcus gattii outbreak. Nature 437: 1360–1364.
26. Byrnes EJ, 3rdBildfell RJ, Frank SA, Mitchell TG, Marr KA, et al. (2009) Molecular evidence that the range of the Vancouver Island outbreak of Cryptococcus gattii infection has expanded into the Pacific Northwest in the United States. J Infect Dis 199: 1081–1086.
27. Hagen F, Ceresini PC, Polacheck I, Ma H, van Nieuwerburgh F, et al. (2013) Ancient dispersal of the human fungal pathogen Cryptococcus gattii from the Amazon rainforest. PLOS ONE 8: e71148.
28. Bovers M, Hagen F, Kuramae EE, Boekhout T, (2008) Six monophyletic lineages identified within Cryptococcus neoformans and Cryptococcus gattii by multi-locus sequence typing. Fungal Genet Biol 45: 400–421.
29. Sharpton TJ, Neafsey DE, Galagan JE, Taylor JW, (2008) Mechanisms of intron gain and loss in Cryptococcus. Genome Biol 9: R24.
30. Xu J, Vilgalys R, Mitchell TG, (2000) Multiple gene genealogies reveal recent dispersion and hybridization in the human pathogenic fungus Cryptococcus neoformans. Mol Ecol 9: 1471–1481.
31. Xu J, Yan Z, Guo H, (2009) Divergence, hybridization, and recombination in the mitochondrial genome of the human pathogenic yeast Cryptococcus gattii. Mol Ecol 18: 2628–2642.
32. Bovers M, Hagen F, Kuramae EE, Boekhout T, (2009) Promiscuous mitochondria in Cryptococcus gattii. FEMS Yeast Res 9: 489–503.
33. Voelz K, Ma H, Phadke S, Byrnes EJ, Zhu P, et al. (2013) Transmission of hypervirulence traits via sexual reproduction within and between lineages of the human fungal pathogen Cryptococcus gattii. PLOS Genetics 9: e1003771.
34. Ma H, Hagen F, Stekel DJ, Johnston SA, Sionov E, et al. (2009) The fatal fungal outbreak on Vancouver Island is characterized by enhanced intracellular parasitism driven by mitochondrial regulation. Proc Natl Acad Sci U S A 106: 12980–12985.
35. Olson A, Stenlid J, (2001) Plant pathogens. Mitochondrial control of fungal hybrid virulence. Nature 411: 438.
36. Byrnes EJ, 3rdBildfell RJ, Dearing PL, Valentine BA, Heitman J, (2009) Cryptococcus gattii with bimorphic colony types in a dog in western Oregon: additional evidence for expansion of the Vancouver Island outbreak. J Vet Diagn Invest 21: 133–136.
37. Walraven CJ, Gerstein W, Hardison SE, Wormley F, Lockhart SR, et al. (2011) Fatal disseminated Cryptococcus gattii infection in New Mexico. PLOS ONE 6: e28625.
38. MacDougall L, Kidd SE, Galanis E, Mak S, Leslie MJ, et al. (2007) Spread of Cryptococcus gattii in British Columbia, Canada, and detection in the Pacific Northwest, USA. Emerg Infect Dis 13: 42–50.
39. Upton A, Fraser JA, Kidd SE, Bretz C, Bartlett KH, et al. (2007) First contemporary case of human infection with Cryptococcus gattii in Puget Sound: Evidence for spread of the Vancouver Island outbreak. J Clin Microbiol 45: 3086–3088.
40. Chaturvedi S, Ren P, Narasipura SD, Chaturvedi V, (2005) Selection of optimal host strain for molecular pathogenesis studies on Cryptococcus gattii. Mycopathologia 160: 207–215.
41. Okamoto K, Hatakeyama S, Itoyama S, Nukui Y, Yoshino Y, et al. (2010) Cryptococcus gattii genotype VGIIa infection in man, Japan, 2007. Emerging Infectious Diseases 16: 1155–1157.
42. Ngamskulrungroj P, Serena C, Gilgado F, Malik R, Meyer W, (2011) Global VGIIa isolates are of comparable virulence to the major fatal Cryptococcus gattii Vancouver Island outbreak genotype. Clinical Microbiology and Infection 17: 251–258.
43. Kaocharoen S, Ngamskulrungroj P, Firacative C, Trilles L, Piyabongkarn D, et al. (2013) Molecular epidemiology reveals genetic diversity amongst isolates of the Cryptococcus neoformans/C. gattii species complex in Thailand. PLoS Negl Trop Dis 7: e2297.
44. Chaturvedi S, Dyavaiah M, Larsen RA, Chaturvedi V, (2005) Cryptococcus gattii in AIDS patients, southern California. Emerging Infectious Diseases 11: 1686–1692.
45. Lockhart SR, Iqbal N, Harris JR, Grossman NT, Debess E, et al. (2013) Cryptococcus gattii in the United States: genotypic diversity of human and veterinary isolates. PLOS ONE 8: e74737.
46. Trilles L, Lazera Mdos S, Wanke B, Oliveira RV, Barbosa GG, et al. (2008) Regional pattern of the molecular types of Cryptococcus neoformans and Cryptococcus gattii in Brazil. Mem Inst Oswaldo Cruz 103: 455–462.
47. Firacative C, Torres G, Rodriguez MC, Escandon P, (2011) First environmental isolation of Cryptococcus gattii serotype B, from Cucuta, Colombia. Biomedica 31: 118–123.
48. Choi YH, Ngamskulrungroj P, Varma A, Sionov E, Hwang SM, et al. (2010) Prevalence of the VNIc genotype of Cryptococcus neoformans in non-HIV-associated cryptococcosis in the Republic of Korea. FEMS Yeast Res 10: 769–778.
49. Chowdhary A, Prakash A, Randhawa HS, Kathuria S, Hagen F, et al. (2013) First environmental isolation of Cryptococcus gattii, genotype AFLP5, from India and a global review. Mycoses 56: 222–228.
50. Olivares LR, Martinez KM, Cruz RM, Rivera MA, Meyer W, et al. (2009) Genotyping of Mexican Cryptococcus neoformans and C. gattii isolates by PCR-fingerprinting. Med Mycol 47: 713–721.
51. Grosse P, Tintelnot K, Sollner O, Schmitz B, (2001) Encephalomyelitis due to Cryptococcus neoformans var gattii presenting as spinal tumour: case report and review of the literature. J Neurol Neurosurg Psychiatry 70: 113–116.
52. Byrnes EJ, 3rdMarr KA, (2011) The outbreak of Cryptococcus gattii in western North America: epidemiology and clinical issues. Current Infectious Disease Reports 13: 256–261.
53. Escandon P, Sanchez A, Firacative C, Castaneda E, (2010) Isolation of Cryptococcus gattii molecular type VGIII, from Corymbia ficifolia detritus in Colombia. Med Mycol 48: 675–678.
54. Mazza M, Refojo N, Bosco-Borgeat ME, Taverna CG, Trovero AC, et al. (2013) Cryptococcus gattii in urban trees from cities in North-eastern Argentina. Mycoses
55. Callejas A, Ordonez N, Rodriguez MC, Castaneda E, (1998) First isolation of Cryptococcus neoformans var. gattii, serotype C, from the environment in Colombia. Med Mycol 36: 341–344.
56. Halliday CL, Bui T, Krockenberger M, Malik R, Ellis DH, et al. (1999) Presence of alpha and a mating types in environmental and clinical collections of Cryptococcus neoformans var. gattii strains from Australia. J Clin Microbiol 37: 2920–2926.
57. Saracli MA, Yildiran ST, Sener K, Gonlum A, Doganci L, et al. (2006) Genotyping of Turkish environmental Cryptococcus neoformans var. neoformans isolates by pulsed field gel electrophoresis and mating type. Mycoses 49: 124–129.
58. Montagna MT, (2002) A note on the isolation of Cryptococcus neoformans serotype A MATa strain from the Italian environment. Med Mycol 40: 593–595.
59. Kidd SE, Hagen F, Tscharke RL, Huynh M, Bartlett KH, et al. (2004) A rare genotype of Cryptococcus gattii caused the cryptococcosis outbreak on Vancouver Island (British Columbia, Canada). Proc Natl Acad Sci USA 101: 17258–17263.
60. Kidd SE, Chow Y, Mak S, Bach PJ, Chen H, et al. (2007) Characterization of environmental sources of the human and animal pathogen Cryptococcus gattii in British Columbia, Canada, and the Pacific Northwest of the United States. Applied Environmental Microbiology 73: 1433–1443.
61. Byrnes EJ, III (2010) Examination of the molecular epidemiology, expansion, population structure, and virulence of the emerging fungal pathogen Cryptococcus gattii in the United States [Ph.D.]. Ann Arbor: Duke University. 311 p.
62. Magditch DA, Liu TB, Xue C, Idnurm A, (2012) DNA mutations mediate microevolution between host-adapted forms of the pathogenic fungus Cryptococcus neoformans. PLoS Pathog 8: e1002936.
63. Toffaletti DL, Nielsen K, Dietrich F, Heitman J, Perfect JR, (2004) Cryptococcus neoformans mitochondrial genomes from serotype A and D strains do not influence virulence. Curr Genet 46: 193–204.
64. Zhu P, Zhai B, Lin X, Idnurm A, (2013) Congenic strains for genetic analysis of virulence traits in Cryptococcus gattii. Infect Immun 81: 2616–2625.
65. Yan Z, Xu J, (2003) Mitochondria are inherited from the MATa parent in crosses of the basidiomycete fungus Cryptococcus neoformans. Genetics 163: 1315–1325.
66. Yan Z, Sun S, Shahid M, Xu J, (2007) Environment factors can influence mitochondrial inheritance in the fungus Cryptococcus neoformans. Fungal Genet Biol 44: 315–322.
67. D'Souza CA, Kronstad JW, Taylor G, Warren R, Yuen M, et al. (2011) Genome variation in Cryptococcus gattii, an emerging pathogen of immunocompetent hosts. mBio 2: e00342–00310.
68. Springer DJ, Ren P, Raina R, Dong Y, Behr MJ, et al. (2010) Extracellular fibrils of pathogenic yeast Cryptococcus gattii are important for ecological niche, murine virulence and human neutrophil interactions. PLOS ONE 5: e10978.
69. Kwon-Chung KJ, Bennett JE, (1984) Epidemiologic differences between the two varieties of Cryptococcus neoformans. Am J Epidemiol 120: 123–130.
70. Kwon-Chung KJ, Bennett JE, (1984) High prevalence of Cryptococcus neoformans var. gattii in tropical and subtropical regions. Zentralbl Bakteriol Mikrobiol Hyg [A] 257: 213–218.
71. Pfeiffer T, Ellis D, (1991) Environmental isolation of Cryptococcus neoformans var. gattii from California. J Infect Dis 163: 929–930.
72. Reed KD, Meece JK, Archer JR, Peterson AT, (2008) Ecologic niche modeling of Blastomyces dermatitidis in Wisconsin. PLoS ONE 3: e2034.
73. McCullough MJ, DiSalvo AF, Clemons KV, Park P, Stevens DA, (2000) Molecular epidemiology of Blastomyces dermatitidis. Clin Infect Dis 30: 328–335.
74. Mandel MA, Barker BM, Kroken S, Rounsley SD, Orbach MJ, (2007) Genomic and population analyses of the mating type loci in Coccidioides species reveal evidence for sexual reproduction and gene acquisition. Eukaryot Cell 6: 1189–1199.
75. Byrnes EJ, 3rdLi W, Lewit Y, Perfect JR, Carter DA, et al. (2009) First reported case of Cryptococcus gattii in the Southeastern USA: implications for travel-associated acquisition of an emerging pathogen. PLOS ONE 4: e5851.
76. Sellers B, Hall P, Cine-Gowdie S, Hays AL, Patel K, et al. (2012) Cryptococcus gattii: an emerging fungal pathogen in the Southeastern United States. Am J Med Sci 343: 510–511.
77. Litvintseva AP, Kestenbaum L, Vilgalys R, Mitchell TG, (2005) Comparative analysis of environmental and clinical populations of Cryptococcus neoformans. J Clin Microbiol 43: 556–564.
78. Loperena-Alvarez Y, Ren P, Li X, Schoonmaker-Bopp DJ, Ruiz A, et al. (2010) Genotypic characterization of environmental isolates of Cryptococcus gattii from Puerto Rico. Mycopathologia 170: 279–285.
79. Litvintseva AP, Thakur R, Vilgalys R, Mitchell TG, (2006) Multilocus sequence typing reveals three genetic subpopulations of Cryptococcus neoformans var. grubii (serotype A), including a unique population in Botswana. Genetics 172: 2223–2238.
80. Velagapudi R, Hsueh YP, Geunes-Boyer S, Wright JR, Heitman J, (2009) Spores as infectious propagules of Cryptococcus neoformans. Infection and Immunity 77: 4345–4355.
81. Su C, Evans D, Cole RH, Kissinger JC, Ajioka JW, et al. (2003) Recent expansion of Toxoplasma through enhanced oral transmission. Science 299: 414–416.
82. Wendte JM, Miller MA, Lambourn DM, Magargal SL, Jessup DA, et al. (2010) Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii. PLoS Genet 6: e1001261.
83. Janbon G, Ormerod KL, Paulet D, Byrnes EJ, 3rdYadav V, et al. (2014) Analysis of the genome and transcriptome of Cryptococcus neoformans var. grubii reveals complex RNA expression and microevolution leading to virulence attenuation. PLoS Genet 10: e1004261.
84. Price JR, Golubchik T, Cole K, Wilson DJ, Crook DW, et al. (2014) Whole-genome sequencing shows that patient-to-patient transmission rarely accounts for acquisition of Staphylococcus aureus in an intensive care unit. Clin Infect Dis 58: 609–618.
85. Steenbergen JN, Shuman HA, Casadevall A, (2001) Cryptococcus neoformans interactions with amoebae suggest an explanation for its virulence and intracellular pathogenic strategy in macrophages. Proc Natl Acad Sci U S A 98: 15245–15250.
86. Steenbergen JN, Nosanchuk JD, Malliaris SD, Casadevall A, (2003) Cryptococcus neoformans virulence is enhanced after growth in the genetically malleable host Dictyostelium discoideum. Infect Immun 71: 4862–4872.
87. Mylonakis E, Ausubel FM, Perfect JR, Heitman J, Calderwood SB, (2002) Killing of Caenorhabditis elegans by Cryptococcus neoformans as a model of yeast pathogenesis. Proc Natl Acad Sci U S A 99: 15675–15680.
88. McClelland EE, Perrine WT, Potts WK, Casadevall A, (2005) Relationship of virulence factor expression to evolved virulence in mouse-passaged Cryptococcus neoformans lines. Infect Immun 73: 7047–7050.
89. Currie B, Sanati H, Ibrahim AS, Edwards JE, JrCasadevall A, et al. (1995) Sterol compositions and susceptibilities to amphotericin B of environmental Cryptococcus neoformans isolates are changed by murine passage. Antimicrob Agents Chemother 39: 1934–1937.
90. Franzot SP, Mukherjee J, Cherniak R, Chen L, Hamdan JS, et al. (1998) Microevolution of a standard strain of Cryptococcus neoformans resulting in differences in virulence and other phenotypes. Infect Immun 66: 89–97.
91. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, et al. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731–2739.
92. Clement M, Posada D, Crandall KA, (2000) TCS: a computer program to estimate gene genealogies. Mol Ecol 9: 1657–1659.
93. Tanaka R, Taguchi H, Takeo K, Miyaji M, Nishimura K, (1996) Determination of ploidy in Cryptococcus neoformans by flow cytometry. J Med Vet Mycol 34: 299–301.
94. DePristo MA, Banks E, Poplin R, Garimella KV, Maguire JR, et al. (2011) A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nat Genet 43: 491–498.
95. Li H, Durbin R, (2009) Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25: 1754–1760.
96. Cingolani P, Platts A, Wang le L, Coon M, Nguyen T, et al. (2012) A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3. Fly (Austin) 6: 80–92.
97. Lassmann T, Sonnhammer EL, (2005) Kalign–an accurate and fast multiple sequence alignment algorithm. BMC Bioinformatics 6: 298.