Multilocus Analysis Resolves the European Finch Epidemic Strain of Trichomonas gallinae and Suggests Introgression from Divergent Trichomonads

Abstract In Europe, Trichomonas gallinae recently emerged as a cause of epidemic disease in songbirds. A clonal strain of the parasite, first found in the United Kingdom, has become the predominant strain there and spread to continental Europe. Discriminating this epidemic strain of T. gallinae from other strains necessitated development of multilocus sequence typing (MLST). Development of the MLST was facilitated by the assembly and annotation of a 54.7 Mb draft genome of a cloned stabilate of the A1 European finch epidemic strain (isolated from Greenfinch, Chloris chloris, XT-1081/07 in 2007) containing 21,924 protein coding genes. This enabled construction of a robust 19 locus MLST based on existing typing loci for Trichomonas vaginalis and T. gallinae. Our MLST has the sensitivity to discriminate strains within existing genotypes confidently, and resolves the American finch A1 genotype from the European finch epidemic A1 genotype. Interestingly, one isolate we obtained from a captive black-naped fruit dove Ptilinopsus melanospilus, was not truly T. gallinae but a hybrid of T. gallinae with a distant trichomonad lineage. Phylogenetic analysis of the individual loci in this fruit dove provides evidence of gene flow between distant trichomonad lineages at 2 of the 19 loci examined and may provide precedence for the emergence of other hybrid trichomonad genomes including T. vaginalis.

[1]  Mike P. Toms,et al.  Health hazards to wild birds and risk factors associated with anthropogenic food provisioning , 2018, Philosophical Transactions of the Royal Society B: Biological Sciences.

[2]  Daniel Mapleson,et al.  KAT: a K-mer analysis toolkit to quality control NGS datasets and genome assemblies , 2016, bioRxiv.

[3]  S. Greenwood,et al.  Molecular characterization of Trichomonas gallinae isolates recovered from the Canadian Maritime provinces’ wild avifauna reveals the presence of the genotype responsible for the European finch trichomonosis epidemic and additional strains , 2015, Parasitology.

[4]  A. von Haeseler,et al.  IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies , 2014, Molecular biology and evolution.

[5]  F. Beaudeau,et al.  The potential capacity of French wildlife rescue centres for wild bird disease surveillance , 2014, European Journal of Wildlife Research.

[6]  J. Carlton,et al.  What is the importance of zoonotic trichomonads for human health? , 2014, Trends in Parasitology.

[7]  S. Lenaghan,et al.  Trichomonas stableri n. sp., an agent of trichomonosis in Pacific Coast band-tailed pigeons (Patagioenas fasciata monilis) , 2013, International journal for parasitology. Parasites and wildlife.

[8]  B. Lawson,et al.  Multi-locus sequence typing confirms the clonality of Trichomonas gallinae isolates circulating in European finches , 2013, Parasitology.

[9]  D. Bell,et al.  The finch epidemic strain of Trichomonas gallinae is predominant in British non-passerines , 2013, Parasitology.

[10]  D. Aanensen,et al.  Genetic Characterization of Trichomonas vaginalis Isolates by Use of Multilocus Sequence Typing , 2012, Journal of Clinical Microbiology.

[11]  D. Bell,et al.  A clonal strain of Trichomonas gallinae is the aetiologic agent of an emerging avian epidemic disease. , 2011, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[12]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[13]  B. Lawson,et al.  Evidence of Spread of the Emerging Infectious Disease, Finch Trichomonosis, by Migrating birds , 2011, EcoHealth.

[14]  M. Hess,et al.  Molecular analysis of clonal trichomonad isolates indicate the existence of heterogenic species present in different birds and within the same host. , 2010, Veterinary parasitology.

[15]  Mike P. Toms,et al.  Emerging Infectious Disease Leads to Rapid Population Declines of Common British Birds , 2010, PloS one.

[16]  Christine K. Johnson,et al.  Clinical Signs and Histopathologic Findings Associated with a Newly Recognized Protozoal Disease (Trichomonas gallinae) in Free-Ranging House Finches (Carpodacus mexicanus) , 2010, Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians.

[17]  M. Forzán,et al.  Trichomoniasis in finches from the Canadian Maritime provinces--An emerging disease. , 2010, The Canadian veterinary journal = La revue veterinaire canadienne.

[18]  K. Handeland,et al.  First Report of Epizootic Trichomoniasis in Wild Finches (Family Fringillidae) in Southern Fennoscandia , 2010, Avian diseases.

[19]  José Sansano-Maestre,et al.  Prevalence and genotyping of Trichomonas gallinae in pigeons and birds of prey , 2009, Avian pathology : journal of the W.V.P.A.

[20]  R. Bondurant,et al.  Studies of trichomonad protozoa in free ranging songbirds: prevalence of Trichomonas gallinae in house finches (Carpodacus mexicanus) and corvids and a novel trichomonad in mockingbirds (Mimus polyglottos). , 2009, Veterinary parasitology.

[21]  M. Yabsley,et al.  Molecular Characterization of the Trichomonas gallinae Morphologic Complex in the United States , 2008, The Journal of parasitology.

[22]  E. Birney,et al.  Velvet: algorithms for de novo short read assembly using de Bruijn graphs. , 2008, Genome research.

[23]  D. Bell,et al.  Molecular identity and heterogeneity of Trichomonad parasites in a closed avian population. , 2007, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[24]  Richard D. Hayes,et al.  Draft Genome Sequence of the Sexually Transmitted Pathogen Trichomonas vaginalis , 2007, Science.

[25]  D. Huson,et al.  Application of phylogenetic networks in evolutionary studies. , 2006, Molecular biology and evolution.

[26]  B. Lawson,et al.  Necrotic ingluvitis in wild finches , 2005, Veterinary Record.

[27]  W. Hanage,et al.  Using Multilocus Sequence Data To Define the Pneumococcus , 2005, Journal of bacteriology.

[28]  Steven Salzberg,et al.  TigrScan and GlimmerHMM: two open source ab initio eukaryotic gene-finders , 2004, Bioinform..

[29]  S. Bonatto,et al.  Molecular phylogeny of Trichomonadidae family inferred from ITS-1, 5.8S rRNA and ITS-2 sequences. , 2004, International journal for parasitology.

[30]  C. Stoeckert,et al.  OrthoMCL: identification of ortholog groups for eukaryotic genomes. , 2003, Genome research.

[31]  S. Salzberg,et al.  Using MUMmer to Identify Similar Regions in Large Sequence Sets , 2004 .

[32]  M. Huynen,et al.  Multiple origins of hydrogenosomes: functional and phylogenetic evidence from the ADP/ATP carrier of the anaerobic chytrid Neocallimastix sp. , 2002, Molecular microbiology.

[33]  M. Achtman,et al.  Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[34]  R. Felleisen Comparative sequence analysis of 5·8S rRNA genes and internal transcribed spacer (ITS) regions of trichomonadid protozoa , 1997, Parasitology.

[35]  P. Cotgreave Relative importance of avian groups in the diets of British and Irish predators , 1995 .

[36]  M. Nei,et al.  Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. , 1993, Molecular biology and evolution.

[37]  J. Felsenstein CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[38]  L. S. Diamond A comparative study of 28 culture media for Trichomonas gallinae. , 1954, Experimental parasitology.

[39]  J. Carlton,et al.  Microsatellite polymorphism in the sexually transmitted human pathogen Trichomonas vaginalis indicates a genetically diverse parasite. , 2011, Molecular and biochemical parasitology.

[40]  K. Henning,et al.  Epidemic mortality in greenfinches at feeder stations caused by Trichomonas gallinae - a recent problem in Northern Germany. , 2009 .

[41]  M. Tibayrenc Multilocus enzyme electrophoresis for parasites and other pathogens. , 2009, Methods in molecular biology.