Sarcocystis Species Richness in Sheep and Goats from Lithuania
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Microscopic Examination for the Presence of Sarcocysts in Goat Tissues
2.3. Molecular Analysis of Sarcocysts Excised from Goat Tissues
2.4. Acid-Pepsin Digestion of Muscle Samples from Sheep and Goats
2.5. Molecular Detection of Specific Sarcocystis spp. in the Tissue Digests
2.6. Phylogenetic and Statistical Analysis
3. Results
3.1. Morphological Observations and Molecular Identification of Sarcocysts Present in Goat Tissues
3.2. Molecular Identification of Sarcocystis spp. in the Digested Samples of Sheep and Goats Using Species-Specific PCR
3.3. The Detection Rates of Sarcocystis spp. in Sheep Tissues by Direct and Nested PCR
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sarcocystis Species | Primer Name | Orientation | Primer Sequence | Ta (°C) | Fragment Size (bp) | Molecular Target |
---|---|---|---|---|---|---|
S. arieticanis | SF1 1 | Forward | ATGGCGTACAACAATCATAAAGAA | 53 | 913 | cox1 |
SsunR3 2 | Reverse | CCGTTGGWATGGCRATCAT | ||||
Arieticanis7F 3 | Forward | TAATTTCCTCGGTACTGTACTGTTTG | 61 | 290 | ||
Arieticanis7R 3 | Reverse | TACTTACGCATTGCGATATTACG | ||||
S. gigantea | V2gig1 2 | Forward | GCACTTCGAGCATTCTTGG | 57 | 548 | |
V2gig2 4 | Reverse | ATCTACATCCACCGTAGGAACCTTA | ||||
V2gig3 4 | Forward | CAGCAAGTACCAAGTTCTGTACGTC | 62 | 322 | ||
V2gig4 2 | Reverse | GGTGCCGAGTACCGAGATACAT | ||||
S. medusiformis | V2medu1 2 | Forward | TTAATGGCATATCGTACTACCTATTG | 56 | 729 | |
V2medu2 2 | Reverse | CCCATGCATCAACCTCCAG | ||||
V2medu3 2 | Forward | GTATCCTGGGGGCCATTAACTT | 61 | 389 | ||
V2medu4 2 | Reverse | CCAAACCAGTGTTCCGAGTATTG | ||||
S. mihoensis | V2miho1 2 | Forward | ATCTTTACACTGCACGGTTTGTTT | 60 | 844 | |
V2miho2 2 | Reverse | AGTCGTTATGTCGGAAGTCAACAG | ||||
V2miho3 2 | Forward | GATGTTACCTCGGGTAAATGCTCTT | 60 | 526 | ||
V2miho4 2 | Reverse | AAAAACATGTCTAGCTCCTAACACC | ||||
S. tenella | SF1 1 | Forward | ATGGCGTACAACAATCATAAAGAA | 53 | 913 | |
SsunR3 2 | Reverse | CCGTTGGWATGGCRATCAT | ||||
Tenella8F 3 | Forward | ATACCGCTCTACGCTGGATCTAC | 59 | 421 | ||
Tenella8R 3 | Reverse | AACCATCGTACAATCCAAAACTAAA | ||||
S. capracanis | VocaF1 3 | Forward | GTAAACTTCCTGGGTACTGTGCTGT | 60 | 531 | |
VocaR1 3 | Reverse | CCAGTAATCCGCTGTCAAGATAC | ||||
V2ca3 3 | Forward | ATACCGATCTTTACGGGAGCAGTA | 58 | 330 | ||
V2ca4 3 | Reverse | GGTCACCGCAGAGAAGTACGAT | ||||
S. hircicanis | V2hirici1 3 | Forward | CCGTAGATGCCATGGGTACTT | 61 | 868 | |
V2hirici2 3 | Reverse | GTAGATATCCAGTGACGTGGTGAG | ||||
V2hirici3 4 | Forward | GCCTGGGTATTCTAGGACTGAGTAG | 62 | 354 | ||
V2hirici4 4 | Reverse | CGAAAACTGCTCTACCGCTCA | ||||
S. morae | SF1 1 | Forward | ATGGCGTACAACAATCATAAAGAA | 53 | 913 | |
SsunR3 2 | Reverse | CCGTTGGWATGGCRATCAT | ||||
V2mor1 3 | Forward | GTGTGCTTGGATCGGTCAAC | 57 | 332 | ||
V2mor2 3 | Reverse | GCCGAATACCGGCTTACTTC | ||||
S. moulei | V2moul1 3 | Forward | GGAGATTCTTGTTGAGTGGGTCT | 54 | 790 | 28S rRNA |
V2moul2 3 | Reverse | GCAAAGCATAATATTTTCTAACGAT | ||||
V2moul3 4 | Forward | GTGAATGCCCTATGTTGTTGAG | 59 | 496 | ||
V2moul4 4 | Reverse | ATATGTGAGAGTGTAGCCCGAAGA |
Species | Host | Information of Sequences | Sequence Similarity, % | |||
---|---|---|---|---|---|---|
n | Length (bp) | GenBank Acc. No. | Comparing Sequences of the Same Species | Comparing Obtained Sequences with Those of Most Closely Related Species | ||
S. tenella | Sheep | 12 | 373 | OR258580-OR258591 | 96.0–100 | 91.7–93.8 S. capracanis |
S. tenella | Goat | 5 | 373 | OR258594-OR258598 | 95.7–100 | 91.4–93.8 S. capracanis |
S. arieticanis | Sheep | 12 | 241 | OR258599-OR258604, OR258607-OR258612 | 96.9–100 * | 84.7–86.7 S. hircicanis |
S. arieticanis | Goat | 1 | 241 | OR258615 | 97.5–100 * | 85.5–86.3 S. hircicanis |
S. capracanis | Goat | 3 | 284 | OR258616-OR258618 | 97.2–99.7 | 90.4–93.7 S. tenella |
S. capracanis | Sheep | 27 | 284 | OR258621-OR258641, OR258647-OR258652 | 96.8–100 | 90.5–93.0 S. tenella |
S. morae | Sheep | 22 | 292 | OR258655-OR258670, OR258672-OR258677 | 95.6–100 | 82.9–85.3 S. cervicanis |
Sarcocystis sp. OaLT1 | Sheep | 2 | 241 | OR258679-OR258680 | No GenBank records | 80.3–83.8 S. tenella 78.6–81.9 S. capracanis 80.4 S. heydorni |
Species | Type of Sample | |||||
---|---|---|---|---|---|---|
Diaphragm | Heart | Oesophagus | ||||
Nested | Direct | Nested | Direct | Nested | Direct | |
S. arieticanis | 44 (93.6) | 28 (59.6) | 39 (83.0) | 5 (10.6) | 39 (83.0) | 9 (19.1) |
S. tenella | 47 (100) | 18 (38.3) | 45 (95.7) | 18 (38.3) | 40 (85.1) | 16 (34.0) |
S. capracanis | 20 (42.6) | 10 (21.3) | 11 (23.4) | 5 (10.6) | 13 (27.7) | 6 (12.8) |
S. morae | 15 (31.9) | 5 (10.6) | 8 (17.0) | 6 (12.8) | 10 (21.3) | 5 (10.6) |
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Marandykina-Prakienė, A.; Butkauskas, D.; Gudiškis, N.; Juozaitytė-Ngugu, E.; Bagdonaitė, D.L.; Kirjušina, M.; Calero-Bernal, R.; Prakas, P. Sarcocystis Species Richness in Sheep and Goats from Lithuania. Vet. Sci. 2023, 10, 520. https://doi.org/10.3390/vetsci10080520
Marandykina-Prakienė A, Butkauskas D, Gudiškis N, Juozaitytė-Ngugu E, Bagdonaitė DL, Kirjušina M, Calero-Bernal R, Prakas P. Sarcocystis Species Richness in Sheep and Goats from Lithuania. Veterinary Sciences. 2023; 10(8):520. https://doi.org/10.3390/vetsci10080520
Chicago/Turabian StyleMarandykina-Prakienė, Alina, Dalius Butkauskas, Naglis Gudiškis, Evelina Juozaitytė-Ngugu, Dovilė Laisvūnė Bagdonaitė, Muza Kirjušina, Rafael Calero-Bernal, and Petras Prakas. 2023. "Sarcocystis Species Richness in Sheep and Goats from Lithuania" Veterinary Sciences 10, no. 8: 520. https://doi.org/10.3390/vetsci10080520