Abstract
The present study describes two cases of Acanthamoeba infections (keratitis and ascites/peritonitis) in small breed domestic dogs in Tenerife, Canary Islands, Spain. In both cases, amoebic trophozoites were observed under the inverted microscope and isolated from the infected tissues and/or fluids, without detecting the presence of other viral, fungal or bacterial pathogens. Amoebae were isolated using 2 % non-nutrient agar plates and axenified for further biochemical and molecular analyses. Osmotolerance and thermotolerance assays revealed that both isolates were able to grow up to 37 °C and 1 M of mannitol and were thus considered as potentially pathogenic. Moreover, the strains were classified as highly cytotoxic as they cause more than 75 % of toxicity when incubated with two eukaryotic cell lines. In order to classify the strains at the molecular level, the diagnostic fragment 3 (DF3) region of the 18S rDNA of Acanthamoeba was amplified and sequenced, revealing that both isolates belonged to genotype T4. In both cases, owners of the animals did not allow any further studies or follow-up and therefore the current status of these animals is unknown. Furthermore, the isolation of these pathogenic amoebae should raise awareness with the veterinary community locally and worldwide.
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Ayers KM, Billups LH, Garner FM (1972) Acanthamoebiasis in a dog. Vet Pathol 9:221–226
Bauer RW, Harrison LR, Watson CW, Styer EL, Chapman WL Jr (1993) Isolation of Acanthamoeba sp. from a greyhound with pneumonia and granulomatous amoebic encephalitis. J Vet Diagn Invest 5:386–391
Booton GC, Visvesvara GS, Byers TJ, Kelly DJ, Fuerst PA (2005) Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol 43:1689–1693
Brofman PJ, Knostman KAB, DiBartola SP (2003) Granulomatous amebic meningoencephalitis causing the syndrome of inappropriate secretion of antidiuretic hormone in a dog. J Vet Int Med 17:230–234
Carlesso AM, Mentz MB, da Machado MLS, Carvalho A, Nunes TET, Maschio VJ, Rott MB (2014) Characterization of isolates of Acanthamoeba from the nasal mucosa and cutaneous lesions of dogs. Curr Microbiol 68:702–707
Dubey JP, Benson JE, Blakeley KT, Booton GC, Visvesvara GS (2005) Disseminated Acanthamoeba sp. infection in a dog. Vet Parasitol 128:183–187
Finnin PJ, Visvesvara GS, Campbell BE, Fry DR, Gasser RB (2007) Multifocal Balamuthia mandrillaris infection in a dog in Australia. Parasitol Res 100:423–426
Foreman O, Sykes J, Ball L, Yang N, De Cock H (2004) Disseminated Infection with Balamuthia mandrillaris in a Dog. Vet Pathol 41:506–510
Hodge PJ, Kelers K, Gasser RB, Visvesvara GS, Martig S, Long SN (2011) Another case of canine amoebic meningoencephalitis—the challenges of reaching a rapid diagnosis. Parasitol Res 108:1069–1073
Ithoi I, Mahmud R, Abdul Basher MH, Jali A, Abdulsalam AM, Ibrahim J, Mak JW (2013) Acanthamoeba genotype T4 detected in naturally-infected feline corneas found to be in homology with those causing human keratitis. Trop Biomed 30(1):131–140
Kent M, Platt SR, Rech RR, Eagleson JS, Howerth EW, Shoff M, Fuerst PA, Booton G, Visvesvara GS, Schatzberg SJ (2011) Multisystemic infection with an Acanthamoeba sp in a dog. JAVMA 238:1476–1481
Lorenzo-Morales J, Ortega-Rivas A, Foronda P, Martínez E, Valladares B (2005) Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources. Parasitol Res 95(4):273–277
Lorenzo-Morales J, Ortega-Rivas A, Martínez E, Khoubbane M, Artigas P, Periago MV, Foronda P, Abreu-Acosta N, Valladares B, Mas-Coma S (2006) Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Trop 100(1–2):63–69
Lorenzo-Morales J, Coronado-Álvarez N, Martínez-Carretero E, Maciver SK, Valladares B (2007) Detection of four adenovirus serotypes within water-isolated strains of Acanthamoeba in the Canary Islands, Spain. Am J Trop Med Hyg. 77(4):753–756
Lorenzo-Morales J, Martín-Navarro CM, López-Arencibia A, Arnalich-Montiel F, Piñero JE, Valladares B (2013) Acanthamoeba keratitis: an emerging disease gathering importance worldwide? Trends Parasitol 29:181–187
Lorenzo-Morales J, Valladares M, Sancho J, Reyes-Batlle M, Martín-Navarro CM, López-Arencibia A, López-Medina L, Piñero JE, Valladares B (2014) First report of a case of prostatitis due to Acanthamoeba in a dog. Acta Protozool 52:325–329
Maciver SK, Asif M, Simmen MW, Lorenzo-Morales J (2013) A systematic analysis of Acanthamoeba genotype frequency correlated with source and pathogenicity: T4 is confirmed as a pathogen-rich genotype. Eur J Protistol 49(2):217–221
Magnet A, Henriques-Gil N, Galván-Diaz AL, Izquierdo F, Fenoy S, del Aguila C (2014) Novel Acanthamoeba 18S rRNA gene sequence type from an environmental isolate. Parasitol Res 113:2845–2850
Martín-Navarro CM, Lorenzo-Morales J, Cabrera-Serra MG, Rancel F, Coronado-Alvarez NM, Piñero JE, Valladares B (2008) The potential pathogenicity of chlorhexidine-sensitive Acanthamoeba strains isolated from contact lens cases from asymptomatic individuals in Tenerife, Canary Islands, Spain. J Med Microbiol 57:1399–1404
Martín-Navarro CM, Lorenzo-Morales J, Machín RP, López-Arencibia A, Valladares B, Piñero JE (2010) Acanthamoeba spp.: in vitro effects of clinical isolates on murine macrophages, osteosarcoma and HeLa cells. Exp Parasitol 126(1):85–88
Niyyati M, Lorenzo-Morales J, Rahimi F, Motevalli-Haghi A, Martín-Navarro CM, Farnia S, Valladares B, Rezaeian M (2009) Isolation and genotyping of potentially pathogenic Acanthamoeba strains from dust sources in Iran. Trans R Soc Trop Med Hyg 103:425–427
Nuprasert W, Putaporntip C, Pariyakanok L, Jongwutiwes S (2010) Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol 48:4636–4640
Pearce JR, Powell HS, Chandler FW, Visvesvara GS (1985) Amebic meningoencephalitis caused by Acanthamoeba castellanii in a dog. J Am Vet Med Assoc 187:951–952
Qvarnstrom Y, Nerad TA, Visvesvara GS (2013) Characterization of a New Pathogenic Acanthamoeba Species, A. byersi n. sp., Isolated from a Human with Fatal Amoebic Encephalitis. J Eukaryot Microbiol 60:626–633
Reed LT, Miller MA, Visvesvara GS, Gardiner CH, Logan MA, Packer RA (2010) Diagnostic exercise. Cerebral mass in a puppy with respiratory distress and progressive neurologic signs. Vet Pathol 47:1116–1119
Reyes-Batlle M, Todd CD, Martín-Navarro CM, López-Arencibia A, Cabello-Vilchez AM, González AC, Córdoba-Linús E, Lindo JF, Valladares B, Piñero JE, Lorenzo-Morales J (2014) Isolation and characterization of Acanthamoeba strains from soil samples in Gran Canaria, Canary Islands, Spain. Parasitol Res 113(4):1383–1388
Siddiqui R, Khan NA (2012) Biology and pathogenesis of Acanthamoeba. Parasit Vectors 5:6
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739
Valladares M, Reyes-Batlle M, Mora-Peces I, Martín-Navarro CM, López-Arencibia A, Dorta-Gorrín A, Comyn-Afonso E, Martínez-Carretero E, Maciver SK, Piñero JE, Valladares B, Lorenzo-Morales J (2014) A multisystemic Acanthamoeba infection in a dog in Tenerife, Canary Islands, Spain. Vet Parasitol 205:707–711
Acknowledgments
This work was supported by the grants RICET (Project No. RD12/0018/0012 of the programme of Redes Temáticas de Investigación Cooperativa, FIS), Spanish Ministry of Health, Madrid, Spain, the Project PI13/00490 “Protozoosis Emergentes por Amebas de Vida Libre: Aislamiento, Caracterización, Nuevas Aproximaciones Terapéuticas y Traslación Clínica de los Resultados” from the Instituto de Salud Carlos III and Project ref. AGUA3 “Amebas de Vida Libre como Marcadores de Calidad del Agua” from CajaCanarias Fundación. ALA and MRB were funded by “Becas de Investigación Obra Social La Caixa-Fundación Cajacanarias para Postgraduados 2014”. JLM was supported by the Ramón y Cajal Subprogramme from the Spanish Ministry of Economy and Competitivity RYC-2011-08863.
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Communicated by Erko Stackebrandt.
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Valladares, M., Reyes-Batlle, M., Martín-Navarro, C.M. et al. Molecular characterization of Acanthamoeba strains isolated from domestic dogs in Tenerife, Canary Islands, Spain. Arch Microbiol 197, 639–643 (2015). https://doi.org/10.1007/s00203-015-1096-1
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DOI: https://doi.org/10.1007/s00203-015-1096-1