Diagnosis of Aspergillosis in Horses
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design, Participants, Sample Processing
2.1.1. Horse with Invasive Equine Pulmonary Aspergillosis (IPA)
2.1.2. Horses with Equine Asthma (EA)
2.1.3. Healthy Controls
2.1.4. Horses with GPM
2.1.5. Sample Processing, Galactomannan and (1,3)-β-D-glucan Assay
2.2. DNA Extraction, PCR, and Sequencing
2.3. Calibration Standards and Fungal Metabolite Extraction
2.4. Liquid Chromatography and Mass Spectrometry Analysis
2.5. Statistical Analysis
3. Results
3.1. Horse with Invasive Equine Pulmonary Aspergillosis (IPA)
3.2. Comparing Asthmatic and Healthy Equine Populations (non-IPA) to IPA
3.3. Horses with GPM
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Individual Equine Data | Sample | Culture | Microscopy (BSH) | PCR | GM (PI) | BDG (pg/mL) | Gtx (ng/mL) | Fc (ng/mL) | TafC (ng/mL) | Conditions |
---|---|---|---|---|---|---|---|---|---|---|
WB, S, 3 yrs. | BALF | A. fumigatus | Positive | A. fumigatus | 2.56 | NA | 86 | Negative | Negative | IPA, EHV-5, Colitis, Death |
Serum | NA | NA | NA | 0.547 | 1142 | Negative | Negative | Negative | ||
QH, G, 9 yrs. | BALF | Negative | Negative | Negative | 0.1 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.379 | 132 | Negative | Negative | Negative | ||
QH, M, 13 yrs. | BALF | A. fumigatus, A. montevidensis, A. flavus, A. niger | Negative | Negative | 2.12 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 2.358 | 387 | Negative | Negative | Negative | ||
TB, G, 9 yrs. | BALF | A. montevidensis | Negative | Negative | 0.15 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.175 | 0 | Negative | Negative | Negative | ||
Sh, G, 13 yrs. | BALF | A. nidulans, R. stolonifer | Negative | Negative | 0.10 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.431 | 16 | Negative | Negative | Negative | ||
WB, G, 14 yrs. | BALF | Negative | Negative | Negative | 0.1 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.16 | 0 | Negative | Negative | Negative | ||
WB, M, 20 yrs. | BALF | Negative | Negative | Negative | 0.1 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.133 | 0 | Negative | Negative | Negative | ||
KB, G, 13 yrs. | BALF | A. montevidensis, A. chevalieri, A. alternata | Negative | Aspergillus sp. | 3.96 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 2.983 | 1291 | Negative | Negative | Negative | ||
WB, G, 13 yrs. | BALF | Negative | Negative | Negative | 0.22 | NA | Negative | Negative | Negative | Severe EA |
Serum | NA | NA | NA | 0.943 | 204 | Negative | Negative | Negative | ||
WB, G, 13 yrs. | BALF | Negative | Negative | Negative | 1.3 | NA | Negative | Negative | Negative | Moderate EA |
Serum | NA | NA | NA | 0.155 | 103 | Negative | Negative | Negative | ||
L, M, 1 yrs. | BALF | Negative | Negative | Negative | 0.24 | NA | Negative | Negative | Negative | Moderate EA |
Serum | NA | NA | NA | 0.216 | 244 | Negative | Negative | Negative | ||
TB, M, 14 yrs. | BALF | A. montevidensis, A. fumigatus, A. alternata | Negative | Negative | 0.86 | NA | Negative | Negative | Negative | Moderate EA |
Serum | NA | NA | NA | 0.65 | 110 | Negative | Negative | Negative | ||
WB, G, 8 yrs. | BALF | A. alternata | Negative | Negative | 0.2 | NA | Negative | Negative | Negative | Moderate EA |
Serum | NA | NA | NA | 0.276 | 139 | Negative | Negative | Negative | ||
CrB, G, 3 yrs. | BALF | Negative | Negative | Negative | 0.31 | NA | Negative | Negative | Negative | Healthy |
Serum | NA | NA | NA | 0.342 | 248 | Negative | Negative | Negative | ||
WB, M, 3 yrs | BALF | Negative | Negative | Negative | 1.48 | NA | Negative | Negative | Negative | Healthy |
Serum | NA | NA | NA | 0.371 | 192 | Negative | Negative | Negative | ||
WB, G, 22 yrs. | BALF | Negative | Negative | Negative | 0.18 | NA | Negative | Negative | Negative | Healthy |
Serum | NA | NA | NA | 0.268 | 142 | Negative | Negative | Negative | ||
WB, M, 13 yrs. | BALF | Negative | Negative | Negative | 0.11 | NA | Negative | Negative | Negative | Healthy |
Serum | NA | NA | NA | 0.257 | 100 | Negative | Negative | Negative | ||
WB, M, 9 yrs. | BALF | Negative | Negative | Negative | 1.5 | NA | Negative | Negative | Negative | Healthy |
Serum | NA | NA | NA | 0.411 | 255 | Negative | Negative | Negative | ||
WB, M, 11 yrs. | Serum | NA | NA | NA | 0.621 | 9 | Negative | Negative | Negative | Healthy |
P, G, 2 yrs. | Serum | NA | NA | NA | 2.629 | 43 | Negative | Negative | Negative | Healthy |
WB, G, 25 yrs. | Serum | NA | NA | NA | 0.5 | 131 | Negative | Negative | Negative | Healthy |
CB, M, 21 yrs. | Serum | NA | NA | NA | 0.606 | 397 | Negative | Negative | Negative | Healthy |
TB, M, 7 yrs. | Serum | NA | NA | NA | 0.257 | 13 | Negative | Negative | Negative | Healthy |
TB, G, 6 yrs. | Serum | NA | NA | NA | 0.105 | 88 | Negative | Negative | Negative | Healthy |
bGM (PI = 2.56) | bAspergillus-DNA | bGtx (86 ng/mL) | sGM (PI = 0.409) | sBDG (1142 pg/mL) | |
---|---|---|---|---|---|
Sensitivity (95% CI) | 100% (0.02–1) | 100% (0.02–1) | 100% (0.02–1) | 100% (0.16–1) | 100% (0.16–1) |
Specificity (95% CI) | 94% (0.71–1) | 88% (0.64–0.99) | 100% (0.8–1) | 55% (0.32–0.77) | 92% (0.62–1) |
PPV | 18% (0.01–0.99) | 33% (0.01–0.91) | 100% (0.02–1) | 18% (0.02–0.52) | 67% (0.09–0.99) |
NPV | 100% (0.79–1) | 100% (0.78–1) | 100% (0.8–1) | 100% (0.72–1) | 92% (0.72–1) |
AUC | 0.941 | 0.941 | 1 | 0.625 | 0.95 |
Tool | Biomarker | ||||||
---|---|---|---|---|---|---|---|
GP | Day | Culture | Aspergillus-DNA | Microscopy (BSH) | GM (PI ≥ 0.5) | Fc (ng/mL) | Sampling |
GPM1 associated with nasopharyngeal dysphagia | |||||||
Left | 1 | A. fumigatus | A. fumigatus | Positive | 9.72 | 492 | BAT |
Left | 7 | A. nidulans | Aspergillus spp. | Positive | 10.4 | 1364 | AS |
GPM2 associated with oral dysphagia | |||||||
Left | 1 | A. nidulans | Aspergillus spp. | Positive | 8.64 | 597 | BAT |
Right | 1 | A. nidulans | Aspergillus spp. | Positive | 8.63 | 3574 | BAT |
Left | 15 | A. nidulans | Negative | Negative | 7.54 | 496 | AS |
Right | 15 | Negative | Negative | Negative | 7.65 | ND | AS |
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Dobiáš, R.; Jahn, P.; Tóthová, K.; Dobešová, O.; Višňovská, D.; Patil, R.; Škríba, A.; Jaworská, P.; Škorič, M.; Podojil, L.; et al. Diagnosis of Aspergillosis in Horses. J. Fungi 2023, 9, 161. https://doi.org/10.3390/jof9020161
Dobiáš R, Jahn P, Tóthová K, Dobešová O, Višňovská D, Patil R, Škríba A, Jaworská P, Škorič M, Podojil L, et al. Diagnosis of Aspergillosis in Horses. Journal of Fungi. 2023; 9(2):161. https://doi.org/10.3390/jof9020161
Chicago/Turabian StyleDobiáš, Radim, Petr Jahn, Katarina Tóthová, Olga Dobešová, Denisa Višňovská, Rutuja Patil, Anton Škríba, Pavla Jaworská, Miša Škorič, Libor Podojil, and et al. 2023. "Diagnosis of Aspergillosis in Horses" Journal of Fungi 9, no. 2: 161. https://doi.org/10.3390/jof9020161