Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM
Background and objectives: Feline leishmaniasis (FeL) is caused by several species of parasites of the genus <i>Leishmania</i>. The disease can occur with the presence or absence of clinical signs, similar to those observed in other common infectious diseases. In endemic regions for FeL,...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/13/15/2468 |
| _version_ | 1827731497246785536 |
|---|---|
| author | Maria Fernanda Alves-Martin Thainá Valente Bertozzo Isabella Neves Aires Suzane Manzini Mirian dos Santos Paixão-Marques Lívia Maísa Guiraldi Wesley José dos Santos Gabriela Pacheco Sánchez Vera Cláudia Lorenzetti Magalhães Curci Virgínia Bodelão Richini-Pereira Simone Baldini Lucheis |
| author_facet | Maria Fernanda Alves-Martin Thainá Valente Bertozzo Isabella Neves Aires Suzane Manzini Mirian dos Santos Paixão-Marques Lívia Maísa Guiraldi Wesley José dos Santos Gabriela Pacheco Sánchez Vera Cláudia Lorenzetti Magalhães Curci Virgínia Bodelão Richini-Pereira Simone Baldini Lucheis |
| author_sort | Maria Fernanda Alves-Martin |
| collection | DOAJ |
| description | Background and objectives: Feline leishmaniasis (FeL) is caused by several species of parasites of the genus <i>Leishmania</i>. The disease can occur with the presence or absence of clinical signs, similar to those observed in other common infectious diseases. In endemic regions for FeL, the infection has been associated with dermatological lesions. Therefore, considering the search for less invasive and more effective diagnostic techniques, we aimed to investigate the presence of <i>Leishmania</i> spp. in domestic cats through Polymerase Chain Reaction (PCR) and high-resolution melting (HRM) analyses of conjunctival, oral, and nasal epithelial cells, and we detected the presence of anti-<i>Leishmania</i> IgG antibodies from serological techniques of the Immunofluorescent Antibody Test (IFAT) and ELISA. Methods: The PCR and HRM for detection of <i>Leishmania</i> spp. were performed on 36 samples of epithelial cells from the conjunctiva of male and female cats, collected using sterile swabs. The serological tests IFAT and ELISA were also performed. Results: The prevalence of <i>Leishmania donovani</i> infection was 11.1% (4/36) by PCR assay, and those results were confirmed for <i>Leishmania</i> species using the HRM technique. Twenty-four cats (24/36 = 66.7%) were reactive to the IFAT and twenty-two cats were reactive by the ELISA technique (22/36 = 61.1%). Interpretation and Conclusions: The use of conjunctival swabs was shown to be a non-invasive, practical, and easy-to-perform technique, and in addition to the genetic sequencing and HRM, it was able to identify the parasitic DNA of <i>L. donovani</i> in cats. This technique can be used for screening diagnosis in future epidemiological surveys of FeL and can be used as a complement to clinical and/or serological tests, as well as associating the clinical history of the animal, for the diagnostic conclusion. |
| first_indexed | 2024-03-11T00:32:03Z |
| format | Article |
| id | doaj.art-28fea5c63367487cbae0bceb8857dbec |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-11T00:32:03Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-28fea5c63367487cbae0bceb8857dbec2023-11-18T22:33:27ZengMDPI AGAnimals2076-26152023-07-011315246810.3390/ani13152468Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRMMaria Fernanda Alves-Martin0Thainá Valente Bertozzo1Isabella Neves Aires2Suzane Manzini3Mirian dos Santos Paixão-Marques4Lívia Maísa Guiraldi5Wesley José dos Santos6Gabriela Pacheco Sánchez7Vera Cláudia Lorenzetti Magalhães Curci8Virgínia Bodelão Richini-Pereira9Simone Baldini Lucheis10Department of Biology and Animal Science, School of Engineering, Sao Paulo State University (UNESP), Ilha Solteira 15385-000, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilDepartment of Molecular Biology and Biochemistry, School of Biological Sciences, University of California, Irvine (UCI), Irvine, CA 92697-2525, USABiological Institute, Araçatuba Regional Laboratory, Araçatuba 16050-230, SP, BrazilAdolfo Lutz Institute, Bauru II Regional Laboratory Center, Bauru 17015-110, SP, BrazilDepartment of Tropical Diseases, Botucatu Medical School, Sao Paulo State University (UNESP), Botucatu 18603-560, SP, BrazilBackground and objectives: Feline leishmaniasis (FeL) is caused by several species of parasites of the genus <i>Leishmania</i>. The disease can occur with the presence or absence of clinical signs, similar to those observed in other common infectious diseases. In endemic regions for FeL, the infection has been associated with dermatological lesions. Therefore, considering the search for less invasive and more effective diagnostic techniques, we aimed to investigate the presence of <i>Leishmania</i> spp. in domestic cats through Polymerase Chain Reaction (PCR) and high-resolution melting (HRM) analyses of conjunctival, oral, and nasal epithelial cells, and we detected the presence of anti-<i>Leishmania</i> IgG antibodies from serological techniques of the Immunofluorescent Antibody Test (IFAT) and ELISA. Methods: The PCR and HRM for detection of <i>Leishmania</i> spp. were performed on 36 samples of epithelial cells from the conjunctiva of male and female cats, collected using sterile swabs. The serological tests IFAT and ELISA were also performed. Results: The prevalence of <i>Leishmania donovani</i> infection was 11.1% (4/36) by PCR assay, and those results were confirmed for <i>Leishmania</i> species using the HRM technique. Twenty-four cats (24/36 = 66.7%) were reactive to the IFAT and twenty-two cats were reactive by the ELISA technique (22/36 = 61.1%). Interpretation and Conclusions: The use of conjunctival swabs was shown to be a non-invasive, practical, and easy-to-perform technique, and in addition to the genetic sequencing and HRM, it was able to identify the parasitic DNA of <i>L. donovani</i> in cats. This technique can be used for screening diagnosis in future epidemiological surveys of FeL and can be used as a complement to clinical and/or serological tests, as well as associating the clinical history of the animal, for the diagnostic conclusion.https://www.mdpi.com/2076-2615/13/15/2468catsPCRHRMconjunctival swabsfeline leishmaniasis |
| spellingShingle | Maria Fernanda Alves-Martin Thainá Valente Bertozzo Isabella Neves Aires Suzane Manzini Mirian dos Santos Paixão-Marques Lívia Maísa Guiraldi Wesley José dos Santos Gabriela Pacheco Sánchez Vera Cláudia Lorenzetti Magalhães Curci Virgínia Bodelão Richini-Pereira Simone Baldini Lucheis Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM Animals cats PCR HRM conjunctival swabs feline leishmaniasis |
| title | Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM |
| title_full | Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM |
| title_fullStr | Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM |
| title_full_unstemmed | Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM |
| title_short | Detection of <i>Leishmania</i> spp. in Cats: Analysis of Nasal, Oral and Conjunctival Swabs by PCR and HRM |
| title_sort | detection of i leishmania i spp in cats analysis of nasal oral and conjunctival swabs by pcr and hrm |
| topic | cats PCR HRM conjunctival swabs feline leishmaniasis |
| url | https://www.mdpi.com/2076-2615/13/15/2468 |
| work_keys_str_mv | AT mariafernandaalvesmartin detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT thainavalentebertozzo detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT isabellanevesaires detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT suzanemanzini detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT miriandossantospaixaomarques detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT liviamaisaguiraldi detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT wesleyjosedossantos detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT gabrielapachecosanchez detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT veraclaudialorenzettimagalhaescurci detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT virginiabodelaorichinipereira detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm AT simonebaldinilucheis detectionofileishmaniaisppincatsanalysisofnasaloralandconjunctivalswabsbypcrandhrm |