Antimicrobial Resistance Profile, Whole-Genome Sequencing and Core Genome Multilocus Sequence Typing of Isolates in Croatia from 2001 to 2022.

Gordan Kompes, Sanja Duvnjak, Irena Reil, ��eljko Mihaljevi��, Boris Habrun, Miroslav Beni��, Luka Cvetni��, Silvio ��pi��i��, Antonela Bagari��
Author Information
  1. Gordan Kompes: Laboratory for General Bacteriology and Mycology, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  2. Sanja Duvnjak: Laboratory for Bacterial Zoonoses and Molecular Diagnostics of Bacterial Diseases, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  3. Irena Reil: Laboratory for Bacterial Zoonoses and Molecular Diagnostics of Bacterial Diseases, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  4. ��eljko Mihaljevi��: Laboratory for Pathology, Department for Pathology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  5. Boris Habrun: Laboratory for General Bacteriology and Mycology, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  6. Miroslav Beni��: Laboratory for Mastitis and Raw Milk Quality, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia.
  7. Luka Cvetni��: Laboratory for Mastitis and Raw Milk Quality, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia.
  8. Silvio ��pi��i��: Laboratory for Bacterial Zoonoses and Molecular Diagnostics of Bacterial Diseases, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia. ORCID
  9. Antonela Bagari��: Laboratory for General Bacteriology and Mycology, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, 10000 Zagreb, Croatia.

Abstract

, the causative agent of anthrax disease, is a worldwide threat to livestock, wildlife and public health. It is also considered one of the most important pathogens of bioterrorism. Rapid and reliable diagnosis and administration of antimicrobials are essential for effective anthrax treatment. In this study, we determined the in vitro susceptibilities of 40 isolates of isolated in Croatia over the recent two decades to 18 antimicrobials. Whole-genome sequencing was performed, and bioinformatics tools were used to determine virulence factors and antimicrobial resistance genes. Core genome-based multilocus sequence typing was used for isolate comparison and phylogenetic analysis. All isolates were susceptible to all antimicrobials recommended for post-exposure prophylaxis or anthrax therapy. Susceptibility was found to all other tested antimicrobials that are an alternative for primary therapy. We found two beta-lactamase genes, but their expression is not sufficient to confer resistance. In all isolates used in this study, we found 21 virulence genes, 8 of which are responsible for toxin and capsule production. As far as phylogenetic analysis is concerned, the isolates from Croatia are categorised into two clades. The first is clade A, subclade Trans Eurasia, and the other is clade B, subclade B2.

Keywords

References

  1. Int J Syst Bacteriol. 1994 Jan;44(1):99-105 [PMID: 8123566]
  2. J Bacteriol. 1996 Jan;178(2):377-84 [PMID: 8550456]
  3. Bioinformatics. 2013 Apr 15;29(8):1072-5 [PMID: 23422339]
  4. Lancet. 2000 Nov 4;356(9241):1574-5 [PMID: 11075776]
  5. BMC Genomics. 2014 Apr 16;15:288 [PMID: 24734872]
  6. Mol Microbiol. 1999 Jun;32(5):1043-53 [PMID: 10361306]
  7. Appl Environ Microbiol. 2019 Mar 6;85(6): [PMID: 30658980]
  8. J Clin Microbiol. 2021 Jun 18;59(7):e0288920 [PMID: 33827898]
  9. J Bacteriol. 2007 Jan;189(1):52-64 [PMID: 17041058]
  10. Antimicrob Agents Chemother. 2002 Jul;46(7):2307-9 [PMID: 12069996]
  11. J Appl Bacteriol. 1993 Nov;75(5):463-72 [PMID: 8300447]
  12. Cent Afr J Med. 1980 Jun;26(6):126-9 [PMID: 7407891]
  13. Antimicrob Agents Chemother. 2003 Jun;47(6):2040-2 [PMID: 12760895]
  14. J Bacteriol. 1964 Jan;87:237-8 [PMID: 14102868]
  15. Sci Rep. 2015 Sep 16;5:14082 [PMID: 26373441]
  16. Toxins (Basel). 2016 Nov 05;8(11): [PMID: 27827957]
  17. Eur J Clin Microbiol Infect Dis. 2011 Jul;30(7):923-30 [PMID: 21279731]
  18. J Clin Microbiol. 2006 Sep;44(9):3422-5 [PMID: 16954291]
  19. Clin Microbiol Infect. 2003 Sep;9(9):984-6 [PMID: 14616693]
  20. Cell Microbiol. 2005 Sep;7(9):1357-64 [PMID: 16098222]
  21. J Clin Microbiol. 2011 Dec;49(12):4286-92 [PMID: 21998431]
  22. Bioinformatics. 2014 Aug 1;30(15):2114-20 [PMID: 24695404]
  23. Antimicrob Agents Chemother. 1998 Dec;42(12):3251-5 [PMID: 9835522]
  24. Antimicrob Agents Chemother. 2010 Oct;54(10):4471-3 [PMID: 20643899]
  25. J Biol Chem. 2009 Jun 5;284(23):15353-7 [PMID: 19201757]
  26. Annu Rev Microbiol. 2009;63:451-76 [PMID: 19514852]
  27. J Bacteriol. 1993 Sep;175(17):5329-38 [PMID: 8366021]
  28. Nat Biotechnol. 2013 Apr;31(4):294-6 [PMID: 23563421]
  29. MMWR Recomm Rep. 2023 Nov 17;72(6):1-47 [PMID: 37963097]
  30. J Clin Microbiol. 2000 Oct;38(10):3780-4 [PMID: 11015402]
  31. BMC Genomics. 2005 Jul 26;6:103 [PMID: 16042811]
  32. J Antimicrob Chemother. 2021 Jan 1;76(1):101-109 [PMID: 33009809]
  33. PLoS One. 2012;7(6):e38054 [PMID: 22701599]
  34. Onderstepoort J Vet Res. 1991 Mar;58(1):17-9 [PMID: 1905000]
  35. J Bacteriol. 2003 Feb;185(3):823-30 [PMID: 12533457]
  36. N Engl J Med. 2001 Nov 29;345(22):1621-6 [PMID: 11704686]
  37. J Clin Microbiol. 2002 Dec;40(12):4732-4 [PMID: 12454180]
  38. Proteomics. 2002 Jun;2(6):784-91 [PMID: 12112862]
  39. Euro Surveill. 2015 Feb 19;20(7):2-4 [PMID: 25719961]
  40. Clin Microbiol Infect. 2017 Jan;23(1):52-54 [PMID: 26794029]
  41. Annu Rev Biochem. 2007;76:243-65 [PMID: 17335404]
  42. Mol Microbiol. 2006 Jun;60(5):1091-8 [PMID: 16689787]
  43. Microbiol Res. 1995 May;150(2):179-86 [PMID: 7600011]
  44. Euro Surveill. 2019 Jun;24(24): [PMID: 31213220]
  45. Biosecur Bioterror. 2013 Sep;11 Suppl 1:S166-76 [PMID: 23971802]
  46. FEMS Microbiol Lett. 2004 Oct 15;239(2):235-40 [PMID: 15476971]
  47. J Bacteriol. 1994 Feb;176(3):586-95 [PMID: 8300513]
  48. BMC Res Notes. 2011 Jul 29;4:273 [PMID: 21801397]
  49. PLoS One. 2020 Jan 13;15(1):e0227875 [PMID: 31931511]
  50. J Antimicrob Chemother. 2004 Feb;53(2):247-51 [PMID: 14688054]
  51. Infect Immun. 1985 Aug;49(2):291-7 [PMID: 3926644]
  52. mSphere. 2019 Aug 7;4(4): [PMID: 31391284]
  53. BMC Microbiol. 2009 Apr 15;9:71 [PMID: 19368722]
  54. Nature. 2003 May 1;423(6935):81-6 [PMID: 12721629]
  55. Br Med J. 1976 Mar 27;1(6012):748 [PMID: 1260311]
  56. J Bacteriol. 2009 Nov;191(21):6683-93 [PMID: 19717606]
  57. mBio. 2016 Sep 27;7(5): [PMID: 27677796]
  58. Microorganisms. 2020 Jan 08;8(1): [PMID: 31936409]
  59. J Gen Microbiol. 1993 Mar;139(3):601-7 [PMID: 8473865]
  60. Clin Infect Dis. 2022 Oct 17;75(Suppl 3):S373-S378 [PMID: 36251548]
  61. J Comput Biol. 2012 May;19(5):455-77 [PMID: 22506599]
  62. Res Microbiol. 2023 Jul-Aug;174(6):104050 [PMID: 36893969]
  63. Appl Environ Microbiol. 1997 Mar;63(3):1054-7 [PMID: 9055420]
  64. J Med Microbiol. 2009 Jun;58(Pt 6):737-744 [PMID: 19429749]
  65. J Med Assoc Thai. 2008 Sep;91(9):1425-32 [PMID: 18843874]
  66. J Antimicrob Chemother. 2007 Sep;60(3):555-67 [PMID: 17586563]
  67. Cell Microbiol. 2012 Aug;14(8):1219-30 [PMID: 22452315]
  68. Ann Inst Pasteur (Paris). 1954 Aug;87(2):215-7 [PMID: 13197885]
  69. Nucleic Acids Res. 2016 Jan 4;44(D1):D694-7 [PMID: 26578559]
  70. Clin Infect Dis. 2007 Jan 15;44(2):280-6 [PMID: 17173232]
  71. Front Vet Sci. 2021 Jul 09;8:674515 [PMID: 34307524]
  72. J Bacteriol. 2004 Jun;186(11):3531-8 [PMID: 15150241]
  73. J Appl Microbiol. 1999 Aug;87(2):261-2 [PMID: 10475962]
  74. Infect Immun. 1983 Jan;39(1):371-6 [PMID: 6401695]
  75. Antimicrob Agents Chemother. 2019 Oct 22;63(11): [PMID: 31427293]
  76. Microb Genom. 2021 Aug;7(8): [PMID: 34402777]
  77. J Bacteriol. 2000 May;182(10):2928-36 [PMID: 10781564]
  78. PLoS One. 2007 May 23;2(5):e461 [PMID: 17520020]
  79. J Bacteriol. 2006 Aug;188(15):5333-44 [PMID: 16855222]
  80. J Biol Chem. 2006 Oct 20;281(42):31408-18 [PMID: 16926147]
  81. J Vet Sci. 2019 Jan 31;20(1):58-62 [PMID: 30541185]
  82. Antibiot Khimioter. 1993 Aug-Sep;38(8-9):34-8 [PMID: 8037574]
  83. Clin Microbiol Infect. 2002 Aug;8(8):479-88 [PMID: 12197870]
  84. Emerg Infect Dis. 2001 Nov-Dec;7(6):933-44 [PMID: 11747719]
  85. Microbiology (Reading). 2010 Oct;156(Pt 10):2982-2993 [PMID: 20688829]
  86. Lancet Infect Dis. 2005 May;5(5):287-95 [PMID: 15854884]
  87. PLoS One. 2014 Jun 05;9(6):e95131 [PMID: 24901417]
  88. Arch Microbiol. 2017 Jul;199(5):775-785 [PMID: 28283680]
  89. Mol Aspects Med. 2009 Dec;30(6):397-405 [PMID: 19729033]

Word Cloud

Created with Highcharts 10.0.0antimicrobialsisolatesanthraxCroatiatwousedresistancegenesfoundstudyvirulenceantimicrobialCorephylogeneticanalysistherapycladesubcladeBcausativeagentdiseaseworldwidethreatlivestockwildlifepublichealthalsoconsideredoneimportantpathogensbioterrorismRapidreliablediagnosisadministrationessentialeffectivetreatmentdeterminedvitrosusceptibilities40isolatedrecentdecades18Whole-genomesequencingperformedbioinformaticstoolsdeterminefactorsgenome-basedmultilocussequencetypingisolatecomparisonsusceptiblerecommendedpost-exposureprophylaxisSusceptibilitytestedalternativeprimarybeta-lactamaseexpressionsufficientconfer218responsibletoxincapsuleproductionfarconcernedcategorisedcladesfirstTransEurasiaB2AntimicrobialResistanceProfileWhole-GenomeSequencingGenomeMultilocusSequenceTypingIsolates20012022anthracisWGScgMLST

Similar Articles

Cited By (1)