Molecular Detection and Characterization of Blastocystis sp. and Enterocytozoon bieneusi in Cattle in Northern Spain
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sampling
2.2. DNA Extraction and Purification
2.3. Molecular Detection of Blastocystis sp.
2.4. Molecular Detection of Enterocytozoon bieneusi
2.5. Sanger Sequencing and Sequence Analysis
2.6. Blastocystis Subtype Identification Using Next Generation Amplicon Sequencing
2.7. Statistical Analysis
3. Results
3.1. Detection of Blastocystis sp. and E. bieneusi by PCR
3.2. Molecular Characterization of Blastocystis sp. and E. bieneusi by Sanger Sequencing
3.3. Blastocystis Subtype Identification by Next Generation Amplicon Sequencing
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Blastocystis sp. | Enterocytozoon bieneusi | ||||
---|---|---|---|---|---|
Variable | n | Positive (n) | % | Positive (n) | % |
Production system | |||||
Dairy | 168 | 38 | 22.6 | 2 | 1.2 |
Beef | 65 | 21 | 32.3 | 0 | 0.0 |
Unknown | 103 | 49 | 47.6 | 0 | 0.0 |
County | |||||
Añana | 11 | 6 | 54.5 | 0 | 0.0 |
Ayala | 106 | 20 | 18.9 | 0 | 0.0 |
Llanada Alavesa | 1 | 1 | 100 | 0 | 0.0 |
Zuia-Gorbeialdea | 118 | 32 | 27.1 | 2 | 1.7 |
Unknown | 100 | 49 | 49.0 | 0 | 0.0 |
Total | 336 | 108 | 32.1 | 2 | 0.6 |
Blastocystis sp. | Enterocytozoon bieneusi | |||
---|---|---|---|---|
Variable | Isolates(n) | Subtypes (n) | Isolates(n) | Genotypes (n) |
Production system | ||||
Dairy | 11 | ST5 (4), ST10 (7) | 2 | BEB4 (2) |
Beef | 9 | ST5 (4), ST10 (5) | 0 | – |
Unknown | 17 | ST5 (9), ST10 (7), ST14 (1) | 0 | – |
Total | 37 | 2 |
Subtype | Next Generation Amplicon Sequencing | Sanger Sequencing |
---|---|---|
ST1 | 1.9% (2/108) | Not detected |
ST3 | 0.9% (1/108) | Not detected |
ST5 | 13.0% (14/108) | 45.9% (17/37) |
ST10 1 | 98.2% (106/108) | 51.4% (19/37) |
ST14 1 | 63.0% (68/108) | 2.7% (1/37) |
ST21 | 64.8% (70/108) | Not detected |
ST23 | 35.2% (38/108) | Not detected |
ST24 | 25.9% (28/108) | Not detected |
ST25 | 86.1% (93/108) | Not detected |
ST26 | 93.5% (101/108) | Not detected |
Number of Subtypes in Co-Infection | Subtype Combinations | Samples (n) |
---|---|---|
2 | ST10 + ST25 | 1 |
ST25 + ST26 | 1 | |
3 | ST5 + ST10 + ST25 | 1 |
ST5 + ST10 + ST14 | 1 | |
ST10 + ST14 + ST24 | 2 | |
ST10 + ST21 + ST26 | 1 | |
ST10 + ST25 + ST26 | 15 | |
ST14 + ST25 + ST26 | 1 | |
4 | ST5 + ST10 + ST25 + ST26 | 3 |
ST10 + ST14 + ST21 + ST24 | 1 | |
ST10 + ST14 + ST25 + ST26 | 5 | |
ST10 + ST21 + ST23 + ST26 | 1 | |
ST10 + ST21 + ST24 + ST26 | 1 | |
ST10 + ST21 + ST25 + ST26 | 4 | |
ST10 + ST23 + ST25 + ST26 | 4 | |
5 | ST5 + ST10 + ST14 + ST25 + ST26 | 1 |
ST5 + ST10 + ST14 + ST24 + ST26 | 1 | |
ST10 + ST14 + ST21 + ST24 + ST26 | 4 | |
ST10 + ST14 + ST21 + ST25 + ST26 | 15 | |
ST10 + ST21 + ST23 + ST25 + ST26 | 8 | |
ST10 + ST14 + ST24 + ST25 + ST26 | 1 | |
6 | ST5 + ST10 + ST14 + ST21 + ST25 + ST26 | 4 |
ST5 + ST10 + ST14 + ST23 + ST25 + ST26 | 1 | |
ST10 + ST14 + ST21 + ST23 + ST24 + ST25 | 1 | |
ST10 + ST14 + ST21 + ST23 + ST24 + ST26 | 3 | |
ST10 + ST14 + ST21 + ST23 + ST25 + ST26 | 12 | |
ST10 + ST14 + ST21 + ST24 + ST25 + ST26 | 5 | |
ST10 + ST21 + ST23 + ST24 + ST25 + ST26 | 1 | |
7 | ST1 + ST5 + ST10 + ST14 + ST21 + ST25 + ST26 | 1 |
ST10 + ST14 + ST21 + ST23 + ST24 + ST25 + ST26 | 7 | |
8 | ST3 + ST10 + ST14 + ST21 + ST23 + ST24 + ST25 + ST26 | 1 |
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Abarca, N.; Santín, M.; Ortega, S.; Maloney, J.G.; George, N.S.; Molokin, A.; Cardona, G.A.; Dashti, A.; Köster, P.C.; Bailo, B.; et al. Molecular Detection and Characterization of Blastocystis sp. and Enterocytozoon bieneusi in Cattle in Northern Spain. Vet. Sci. 2021, 8, 191. https://doi.org/10.3390/vetsci8090191
Abarca N, Santín M, Ortega S, Maloney JG, George NS, Molokin A, Cardona GA, Dashti A, Köster PC, Bailo B, et al. Molecular Detection and Characterization of Blastocystis sp. and Enterocytozoon bieneusi in Cattle in Northern Spain. Veterinary Sciences. 2021; 8(9):191. https://doi.org/10.3390/vetsci8090191
Chicago/Turabian StyleAbarca, Nadia, Mónica Santín, Sheila Ortega, Jenny G. Maloney, Nadja S. George, Aleksey Molokin, Guillermo A. Cardona, Alejandro Dashti, Pamela C. Köster, Begoña Bailo, and et al. 2021. "Molecular Detection and Characterization of Blastocystis sp. and Enterocytozoon bieneusi in Cattle in Northern Spain" Veterinary Sciences 8, no. 9: 191. https://doi.org/10.3390/vetsci8090191