Diagnosis of Aspergillosis in Horses.
- Journal Article
Summary
The research article concerns the diagnosis of invasive pulmonary aspergillosis (IPA) in horses, a potentially fatal condition. Using serum samples and bronchoalveolar lavage fluid, the presence of IPA was detected by analyzing differences in metabolite concentrations between healthy horses, those with equine asthma, and those with IPA.
Introduction
In this research paper, the authors aim to explore direct diagnostic tools for detecting invasive pulmonary aspergillosis (IPA) in horses, a fungal infection that causes a severe form of pneumonia. In horses, the condition is rare but deadly, with nearly all cases resulting in mortality. Accurate and early diagnosis is crucial for a chance at successful treatment.
Data Collection and Testing
- The nature of the sample collection included both bronchoalveolar lavage fluid (BALF)— a method to examine the cells and fluid in the air sacs of the lungs— and serum samples acquired from blood tests.
- The samples were collected from 18 horses and categorized into three groups: those displaying signs of invasive pulmonary aspergillosis (IPA), those with a chronic respiratory condition called equine asthma (EA), and 5 healthy control horses. Additional serum samples were collected from another 6 healthy controls.
- Various tests were performed on the samples to detect different fungal elements: spp. DNA, fungal galactomannan (GM), ferricrocin (Fc), triacetylfusarinin C (TafC), and gliotoxin (Gtx).
Analysis and Results
- Analysis of 24 serum samples for (1,3)-β-D-glucan (BDG) and GM was conducted. BDG is a glucose polymer found in fungal cells, and increased levels in serum are commonly associated with fungal infection. The median BDG levels in control horses’ serum were 131 pg/mL, while in IPA diagnosed horses’ serum, this was significantly higher—1142 pg/mL.
- Similar trends were found in BALF samples for GM and DNA. The Area under the Curve (AUC) score of 0.941 signifies a very high level of diagnostic accuracy, suggesting elevated levels of these compounds are strong indicators of IPA.
- Additionally, they detected the fungal secondary metabolite Gtx in the IPA horses’ BALF and lung tissue samples. Gtx is a toxic compound produced by the Aspergillus fungus, and its presence further confirms an IPA infection.
Conclusion
Overall, the study shows that detecting DNA and various fungal metabolites in BALF and serum samples from horses can provide a robust method of diagnosing IPA. This research contributes to the understanding of IPA diagnosis in horses and may improve their prognosis.
Cite This Article
Publication
Researcher Affiliations
- Department of Bacteriology and Mycology, Public Health Institute in Ostrava, 702 00 Ostrava, Czech Republic.
- Department of Biomedical Sciences, Faculty of Medicine, University of Ostrava, 703 00 Ostrava, Czech Republic.
- Equine Clinic, University of Veterinary Sciences, 612 42 Brno, Czech Republic.
- Equine Clinic, University of Veterinary Sciences, 612 42 Brno, Czech Republic.
- Equine Clinic, University of Veterinary Sciences, 612 42 Brno, Czech Republic.
- Department of Biomedical Sciences, Faculty of Medicine, University of Ostrava, 703 00 Ostrava, Czech Republic.
- Department of Biology and Ecology, Faculty of Science, University of Ostrava, 710 00 Ostrava, Czech Republic.
- Institute of Microbiology of the Czech Academy of Sciences, 142 20 Prague, Czech Republic.
- Department of Analytical Chemistry, Faculty of Science, Palacký University, 771 46 Olomouc, Czech Republic.
- Institute of Microbiology of the Czech Academy of Sciences, 142 20 Prague, Czech Republic.
- Department of Bacteriology and Mycology, Public Health Institute in Ostrava, 702 00 Ostrava, Czech Republic.
- Department of Pathology and Parasitology, University of Veterinary Sciences Brno, 612 42 Brno, Czech Republic.
- Equine Clinic, University of Veterinary Sciences, 612 42 Brno, Czech Republic.
- Department of Molecular Biology, Public Health Institute in Ostrava, 702 00 Ostrava, Czech Republic.
- Department of Molecular Biology, Public Health Institute in Ostrava, 702 00 Ostrava, Czech Republic.
- Department of Bacteriology and Mycology, Public Health Institute in Ostrava, 702 00 Ostrava, Czech Republic.
- Department of Biomedical Sciences, Faculty of Medicine, University of Ostrava, 703 00 Ostrava, Czech Republic.
- California Institute for Medical Research, San Jose, CA 95128, USA.
- Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, CA 95128, USA.
- Institute of Microbiology of the Czech Academy of Sciences, 142 20 Prague, Czech Republic.
- Department of Analytical Chemistry, Faculty of Science, Palacký University, 771 46 Olomouc, Czech Republic.
Grant Funding
- 21-17044S / Grant Agency of the Czech Republic
- 109/2019/FVL / Internal Grant Agency of the University of Veterinary Sciences Brno
Conflict of Interest Statement
References
- Seyedmousavi S, Guillot J, Arné P, de Hoog GS, Mouton JW, Melchers WJ, Verweij PE. Aspergillus and aspergilloses in wild and domestic animals: a global health concern with parallels to human disease.. Med Mycol 2015 Nov;53(8):765-97.
- Tell LA. Aspergillosis in mammals and birds: impact on veterinary medicine.. Med Mycol 2005 May;43 Suppl 1:S71-3.
- Latgé JP, Chamilos G. Aspergillus fumigatus and Aspergillosis in 2019.. Clin Microbiol Rev 2019 Dec 18;33(1).
- Cafarchia C, Figueredo LA, Otranto D. Fungal diseases of horses.. Vet Microbiol 2013 Nov 29;167(1-2):215-34.
- Dobesova O, Schwarz B, Velde K, Jahn P, Zert Z, Bezdekova B. Guttural pouch mycosis in horses: a retrospective study of 28 cases.. Vet Rec 2012 Dec 1;171(22):561.
- Cousty M, Tricaud C, De Beauregard T, Picandet V, Bizon-Mercier C, Tessier C. Ligation of the ipsilateral common carotid artery and topical treatment for the prevention of epistaxis from guttural pouch mycosis in horses.. Vet Rec 2016 Jan 9;178(2):44.
- Watkins AR, Parente EJ. Salpingopharyngeal fistula as a treatment for guttural pouch mycosis in seven horses.. Equine Vet J 2018 Nov;50(6):781-786.
- Lepage OM, Di Francesco P, Moulin N, Gangl M, Texier G, Marchi J, Cadoré JL. The Effect of Topical Oxygen Therapy in Horses Affected with Mycosis of the Guttural Pouch: An Experimental Pilot Study and a Case Series.. Animals (Basel) 2021 Nov 22;11(11).
- Pace LW, Wirth NR, Foss RR, Fales WH. Endocarditis and pulmonary aspergillosis in a horse.. J Vet Diagn Invest 1994 Oct;6(4):504-6.
- Johnson PJ, Moore LA, Mrad DR, Turk JR, Wilson DA. Sudden death of two horses associated with pulmonary aspergillosis.. Vet Rec 1999 Jul 3;145(1):16-20.
- Elad D, Segal E. Diagnostic Aspects of Veterinary and Human Aspergillosis.. Front Microbiol 2018;9:1303.
- Khot PD, Ko DL, Fredricks DN. Sequencing and analysis of fungal rRNA operons for development of broad-range fungal PCR assays.. Appl Environ Microbiol 2009 Mar;75(6):1559-65.
- Kumar M, Shukla PK. Use of PCR targeting of internal transcribed spacer regions and single-stranded conformation polymorphism analysis of sequence variation in different regions of rrna genes in fungi for rapid diagnosis of mycotic keratitis.. J Clin Microbiol 2005 Feb;43(2):662-8.
- Škríba A, Patil RH, Hubáček P, Dobiáš R, Palyzová A, Marešová H, Pluháček T, Havlíček V. Rhizoferrin Glycosylation in Rhizopus microsporus.. J Fungi (Basel) 2020 Jun 18;6(2).
- Novák J, Škríba A, Havlíček V. CycloBranch 2: Molecular Formula Annotations Applied to imzML Data Sets in Bimodal Fusion and LC-MS Data Files.. Anal Chem 2020 May 19;92(10):6844-6849.
- R Core Team. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing; Vienna, Austria: 2020.
- Thiele C, Hirschfeld G. Cutpointr: Improved estimation and validation of optimal cutpoints in R. arXiv 2020.
- Stevenson M, Sergeant E, Nunes T, Heuer C, Marshall J, Sanchez J, Thornton R, Reiczigel J, Robison-Cox J, Sebastiani P. epiR: Tools for the Analysis of Epidemiological Data. R Package Version 2.0.33 2021.
- Hattel AL, Drake TR, Anderholm BJ, McAllister ES. Pulmonary aspergillosis associated with acute enteritis in a horse.. J Am Vet Med Assoc 1991 Sep 1;199(5):589-90.
- Sweeney CR, Habecker PL. Pulmonary aspergillosis in horses: 29 cases (1974-1997).. J Am Vet Med Assoc 1999 Mar 15;214(6):808-11.
- Green SL, Hager DA, Calderwood Mays MB, Merritt A. Acute diffuse mycotic pneumonia in a 7-month-old colt. Vet. Radiol. 1987;28:216–219.
- Slocombe RF, Slauson DO. Invasive pulmonary aspergillosis of horses: an association with acute enteritis.. Vet Pathol 1988 Jul;25(4):277-81.
- Rosenstein D, Mullaney T. Acute pulmonary aspergillosis and severe enterocolitis in a quarter horse mare. Equine Vet. Educ. 1996;8:200–203.
- Couëtil LL, Hoffman AM, Hodgson J, Buechner-Maxwell V, Viel L, Wood JL, Lavoie JP. Inflammatory airway disease of horses.. J Vet Intern Med 2007 Mar-Apr;21(2):356-61.
- Couëtil LL, Cardwell JM, Gerber V, Lavoie JP, Léguillette R, Richard EA. Inflammatory Airway Disease of Horses--Revised Consensus Statement.. J Vet Intern Med 2016 Mar-Apr;30(2):503-15.
- Donnelly JP, Chen SC, Kauffman CA, Steinbach WJ, Baddley JW, Verweij PE, Clancy CJ, Wingard JR, Lockhart SR, Groll AH, Sorrell TC, Bassetti M, Akan H, Alexander BD, Andes D, Azoulay E, Bialek R, Bradsher RW, Bretagne S, Calandra T, Caliendo AM, Castagnola E, Cruciani M, Cuenca-Estrella M, Decker CF, Desai SR, Fisher B, Harrison T, Heussel CP, Jensen HE, Kibbler CC, Kontoyiannis DP, Kullberg BJ, Lagrou K, Lamoth F, Lehrnbecher T, Loeffler J, Lortholary O, Maertens J, Marchetti O, Marr KA, Masur H, Meis JF, Morrisey CO, Nucci M, Ostrosky-Zeichner L, Pagano L, Patterson TF, Perfect JR, Racil Z, Roilides E, Ruhnke M, Prokop CS, Shoham S, Slavin MA, Stevens DA, Thompson GR, Vazquez JA, Viscoli C, Walsh TJ, Warris A, Wheat LJ, White PL, Zaoutis TE, Pappas PG. Revision and Update of the Consensus Definitions of Invasive Fungal Disease From the European Organization for Research and Treatment of Cancer and the Mycoses Study Group Education and Research Consortium.. Clin Infect Dis 2020 Sep 12;71(6):1367-1376.
- Douglas AP, Smibert OC, Bajel A, Halliday CL, Lavee O, McMullan B, Yong MK, van Hal SJ, Chen SC. Consensus guidelines for the diagnosis and management of invasive aspergillosis, 2021.. Intern Med J 2021 Nov;51 Suppl 7:143-176.
- Ullmann AJ, Aguado JM, Arikan-Akdagli S, Denning DW, Groll AH, Lagrou K, Lass-Flörl C, Lewis RE, Munoz P, Verweij PE, Warris A, Ader F, Akova M, Arendrup MC, Barnes RA, Beigelman-Aubry C, Blot S, Bouza E, Brüggemann RJM, Buchheidt D, Cadranel J, Castagnola E, Chakrabarti A, Cuenca-Estrella M, Dimopoulos G, Fortun J, Gangneux JP, Garbino J, Heinz WJ, Herbrecht R, Heussel CP, Kibbler CC, Klimko N, Kullberg BJ, Lange C, Lehrnbecher T, Löffler J, Lortholary O, Maertens J, Marchetti O, Meis JF, Pagano L, Ribaud P, Richardson M, Roilides E, Ruhnke M, Sanguinetti M, Sheppard DC, Sinkó J, Skiada A, Vehreschild MJGT, Viscoli C, Cornely OA. Diagnosis and management of Aspergillus diseases: executive summary of the 2017 ESCMID-ECMM-ERS guideline.. Clin Microbiol Infect 2018 May;24 Suppl 1:e1-e38.
- Meersseman W, Lagrou K, Maertens J, Wilmer A, Hermans G, Vanderschueren S, Spriet I, Verbeken E, Van Wijngaerden E. Galactomannan in bronchoalveolar lavage fluid: a tool for diagnosing aspergillosis in intensive care unit patients.. Am J Respir Crit Care Med 2008 Jan 1;177(1):27-34.
- D'Haese J, Theunissen K, Vermeulen E, Schoemans H, De Vlieger G, Lammertijn L, Meersseman P, Meersseman W, Lagrou K, Maertens J. Detection of galactomannan in bronchoalveolar lavage fluid samples of patients at risk for invasive pulmonary aspergillosis: analytical and clinical validity.. J Clin Microbiol 2012 Apr;50(4):1258-63.
- Cao XJ, Li YP, Xie LM, Zhang HL, Qin YS, Guo XG. Diagnostic Accuracy of Bronchoalveolar Lavage Fluid Galactomannan for Invasive Aspergillosis.. Biomed Res Int 2020;2020:5434589.
- Cabana AL, Xavier MO, Mendes JF, Teles AJ, Martins AM, Silva-Filho RP, Meireles MCA. Applicability of the Platelia EIA® Aspergillus test for the diagnosis of aspergilosis in penguins.. Braz J Biol 2019 Apr-Jun;79(2):169-173.
- Garcia RS, Wheat LJ, Cook AK, Kirsch EJ, Sykes JE. Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs.. J Vet Intern Med 2012 Jul-Aug;26(4):911-9.
- Monteiro AA, Rubenich DS, Zandoná MR, Pasqualotto AC. Impact of pre-analytical variables in the determination of serum galactomannan.. Med Mycol 2017 Aug 1;55(6):635-641.
- Dobias R, Jaworska P, Tomaskova H, Kanova M, Lyskova P, Vrba Z, Holub C, Svobodová L, Hamal P, Raska M. Diagnostic value of serum galactomannan, (1,3)-β-d-glucan, and Aspergillus fumigatus-specific IgA and IgG assays for invasive pulmonary aspergillosis in non-neutropenic patients.. Mycoses 2018 Aug;61(8):576-586.
- Dobiáš R, Škríba A, Pluháček T, Petřík M, Palyzová A, Káňová M, Čubová E, Houšť J, Novák J, Stevens DA, Mitulovič G, Krejčí E, Hubáček P, Havlíček V. Noninvasive Combined Diagnosis and Monitoring of Aspergillus and Pseudomonas Infections: Proof of Concept.. J Fungi (Basel) 2021 Sep 6;7(9).
- Vergidis P, Moore CB, Novak-Frazer L, Rautemaa-Richardson R, Walker A, Denning DW, Richardson MD. High-volume culture and quantitative real-time PCR for the detection of Aspergillus in sputum.. Clin Microbiol Infect 2020 Jul;26(7):935-940.
- Levesque E, Rizk F, Noorah Z, Aït-Ammar N, Cordonnier-Jourdin C, El Anbassi S, Bonnal C, Azoulay D, Merle JC, Botterel F. Detection of (1,3)-β-d-Glucan for the Diagnosis of Invasive Fungal Infection in Liver Transplant Recipients.. Int J Mol Sci 2017 Apr 19;18(4).
- White PL, Wiederhold NP, Loeffler J, Najvar LK, Melchers W, Herrera M, Bretagne S, Wickes B, Kirkpatrick WR, Barnes RA, Donnelly JP, Patterson TF. Comparison of Nonculture Blood-Based Tests for Diagnosing Invasive Aspergillosis in an Animal Model.. J Clin Microbiol 2016 Apr;54(4):960-6.
- Owens RA, Doyle S. Effects of antifungal agents on the fungal proteome: informing on mechanisms of sensitivity and resistance.. Expert Rev Proteomics 2021 Mar;18(3):185-199.
- Kriegl L, Havlicek V, Dichtl K, Egger M, Hoenigl M. Siderophores: a potential role as a diagnostic for invasive fungal disease.. Curr Opin Infect Dis 2022 Dec 1;35(6):485-492.
- Dobiáš R, Jaworská P, Skopelidou V, Strakoš J, Višňovská D, Káňová M, Škríba A, Lysková P, Bartek T, Janíčková I, Kozel R, Cwiková L, Vrba Z, Navrátil M, Martinek J, Coufalová P, Krejčí E, Ulmann V, Raška M, Stevens DA, Havlíček V. Distinguishing Invasive from Chronic Pulmonary Infections: Host Pentraxin 3 and Fungal Siderophores in Bronchoalveolar Lavage Fluids.. J Fungi (Basel) 2022 Nov 12;8(11).
- Luptáková D, Patil RH, Dobiáš R, Stevens DA, Pluháček T, Palyzová A, Káňová M, Navrátil M, Vrba Z, Hubáček P, Havlíček V. Siderophore-Based Noninvasive Differentiation of Aspergillus fumigatus Colonization and Invasion in Pulmonary Aspergillosis.. Microbiol Spectr 2023 Jan 31;11(2):e0406822.
Citations
This article has been cited 2 times.- Dobiáš R, Stevens DA, Havlíček V. Current and Future Pathways in Aspergillus Diagnosis. Antibiotics (Basel) 2023 Feb 13;12(2).
- Melo UP, Ferreira C, Barreto SWM. Pulmonary aspergillosis in a horse: a case report. Braz J Vet Med 2024;46:e004723.