- Volume 57 Issue 2
DOI QR Code
Detection and molecular characterization of Hepatozoon canis, Babesia vogeli, Ehrlichia canis, and Anaplasma platys in dogs from Metro Manila, Philippines
- Adao, Davin Edric V. (Institute of Biology, College of Science, Natural Sciences Research Institute, University of the Philippines Diliman) ;
- Herrera, Charles Michael T. (Institute of Biology, College of Science, Natural Sciences Research Institute, University of the Philippines Diliman) ;
- Galarion, Luiza H. (Institute of Biology, College of Science, Natural Sciences Research Institute, University of the Philippines Diliman) ;
- Bolo, Nicole R. (Institute of Biology, College of Science, Natural Sciences Research Institute, University of the Philippines Diliman) ;
- Carlos, Rhodora S. (Carlos Veterinary Clinic) ;
- Carlos, Enrique T. (Makati Dog and Cat Hospital) ;
- Carlos, Sixto S. (Makati Dog and Cat Hospital) ;
- Rivera, Windell L. (Institute of Biology, College of Science, Natural Sciences Research Institute, University of the Philippines Diliman)
- Received : 2017.01.19
- Accepted : 2017.04.21
- Published : 2017.06.30
The study of canine vector-borne diseases in the Philippines started in the 1970s but only gained interest in the past decade. Characterization of such diseases in the Philippines remains incomplete, thus, it is necessary to obtain additional information on the prevalence and diversity of canine tick-borne diseases in the country. In this study, blood samples were obtained at two veterinary clinics in Metro Manila, Philippines from 114 dogs suspected of having canine tick-borne pathogens. Polymerase chain reaction (PCR) was performed on whole blood DNA extracts followed by sequencing, and the following pathogens were detected: Hepatozoon (H.) canis (5.26%), Babesia (B.) vogeli (5.26%), Ehrlichia (E.) canis (4.39%), and Anaplasma platys (3.51%). Additionally, a set of multiplex PCR primers were developed to detect H. canis, Babesia spp. (B. canis and B. vogeli), and E. canis in canine blood. Multiplex and conventional single-reaction PCR results for the 114 dog blood samples were similar, except for one H. canis sample. Multiplex PCR is, therefore, a useful tool in screening infected dogs in veterinary clinics. This study's results, together with those of previous studies in the country, show that canine vector-borne pathogens are an emerging veterinary concern in the Philippines.
Supported by : Natural Sciences Research Institute
- Brecher ME, Hay SN. Bacterial contamination of blood components. Clin Microbiol Rev 2005, 18, 195-204. https://doi.org/10.1128/CMR.18.1.195-204.2005
- Bulla C, Takahira RK, Araujo JP Jr, Aparecida Trinca L, Souza Lopes R, Wiedmeyer CE. The relationship between the degree of thrombocytopenia and infection with Ehrlichia canis in an endemic area. Vet Res 2004, 35, 141-146. https://doi.org/10.1051/vetres:2003038
- Cardoso L, Yisaschar-Mekuzas Y, Rodrigues FT, Costa A, Machado J, Diz-Lopes D, Baneth G. Canine babesiosis in northern Portugal and molecular characterization of vectorborne co-infections. Parasit Vectors 2010, 3, 27. https://doi.org/10.1186/1756-3305-3-27
- Carlos ET, Carlos ER, Calalay FT, Cabiles CC. Babesia canis: clinical observations in dogs in the Philippines. Philipp J Vet Med 1972, 11, 81-91.
- Carlos ET, Cruz FB, Cabiles CC, Calalay FT, Carlos ER. Hepatozoon sp. in the WBC of a human patient. UP Vet 1971, 15, 5-7.
- Corales JMI, Viloria VV, Venturina VM, Mingala CN. The prevalence of Ehrlichia canis, Anaplasma platys and Babesia spp. in dogs in Nueva Ecija, Philippines based on multiplex polymerase chain reaction (mPCR) assay. Ann Parasitol 2014, 60, 267-272.
- Foldvari G, Hell E, Farkas R. Babesia canis canis in dogs from Hungary: detection by PCR and sequencing. Vet Parasitol 2005, 127, 221-226. https://doi.org/10.1016/j.vetpar.2004.10.016
- Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 2010, 59, 307-321. https://doi.org/10.1093/sysbio/syq010
- Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 1999, 41, 95-98.
- Inokuma H, Beppu T, Okuda M, Shimada Y, Sakata Y. Detection of ehrlichial DNA in Haemaphysalis ticks recovered from dogs in Japan that is closely related to a novel Ehrlichia sp. found in cattle ticks from Tibet, Thailand, and Africa. J Clin Microbiol 2004, 42, 1353-1355. https://doi.org/10.1128/JCM.42.3.1353-1355.2004
- Inokuma H, Okuda M, Ohno K, Shimoda K, Onishi T. Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Vet Parasitol 2002, 106, 265-271. https://doi.org/10.1016/S0304-4017(02)00065-1
- Inpankaew T, Hii SF, Chimnoi W, Traub RJ. Canine vector-borne pathogens in semi-domesticated dogs residing in northern Cambodia. Parasit Vectors 2016, 9, 253. https://doi.org/10.1186/s13071-016-1552-z
- Irwin PJ. Canine babesiosis: from molecular taxonomy to control. Parasit Vectors 2009, 2 (Suppl 1), S4.
- Jefferies R, Ryan UM, Muhlnickel CJ, Irwin PJ. Two species of canine Babesia in Australia: detection and characterization by PCR. J Parasitol 2003, 89, 409-412. https://doi.org/10.1645/0022-3395(2003)089[0409:TSOCBI]2.0.CO;2
- Baneth G, Mathew JS, Shkap V, Macintire DK, Barta JR, Ewing SA. Canine hepatozoonosis: two disease syndromes caused by separate Hepatozoon spp. Trends Parasitol 2003, 19, 27-31. https://doi.org/10.1016/S1471-4922(02)00016-8
- Baticados AM, Baticados WN, Carlos ET, Carlos SMEAS, Villarba LA, Subiaga SG, Magcalas JM. Parasitological detection and molecular evidence of Hepatozoon canis from canines in Manila, Philippines. Vet Med Res Rep 2010, 1, 7-10.
- Kelly PJ, Lucas H. Failure to demonstrate Babesia, Anaplasma or Ehrlichia in thrombocytopenic dogs from St Kitts. J Infect Dev Ctries 2009, 3, 561-563.
- Kledmanee K, Suwanpakdee S, Krajangwong S, Chatsiriwech J, Suksai P, Suwannachat P, Sariya L, Buddhirongawatr R, Charoonrut P, Chaichoun K. Development of multiplex polymerase chain reaction for detection of Ehrlichia canis, Babesia spp and Hepatozoon canis in canine blood. Southeast Asian J Trop Med Public Health 2009, 40, 35-39.
- Koh FX, Panchadcharam C, Tay ST. Vector-borne diseases in stray dogs in peninsular Malaysia and molecular detection of Anaplasma and Ehrlichia spp. from Rhipicephalus sanguineus (Acari: Ixodidae) ticks. J Med Entomol 2016, 53, 183-187. https://doi.org/10.1093/jme/tjv153
- Liu M, Ruttayaporn N, Saechan V, Jirapattharasate C, Vudriko P, Moumouni PFA, Cao S, Inpankaew T, Ybanez AP, Suzuki H, Xuan X. Molecular survey of canine vectorborne diseases in stray dogs in Thailand. Parasitol Int 2016, 65, 357-361. https://doi.org/10.1016/j.parint.2016.04.011
- Macieira DB, Messick JB, Cerqueira AMF, Freire IMA, Linhares GFC, Almeida NKO, Almosny NRP. Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil. Vet Clin Pathol 2005, 34, 44-48. https://doi.org/10.1111/j.1939-165X.2005.tb00008.x
- Maggi RG, Birkenheuer AJ, Hegarty BC, Bradley JM, Levy MG, Breitschwerdt EB. Comparison of serological and molecular panels for diagnosis of vector-borne diseases in dogs. Parasit Vectors 2014, 7, 127. https://doi.org/10.1186/1756-3305-7-127
- Mylonakis ME, Koutinas AF, Billinis C, Leontides LS, Kontos V, Papadopoulos O, Rallis T, Fytianou A. Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods. Vet Microbiol 2003, 91, 197-204. https://doi.org/10.1016/S0378-1135(02)00298-5
- Novilla MN, Kwapien RP, Peneyra RS. Occurrence of canine hepatozoonosis in the Philippines. Proc Helminthol Soc Wash 1977, 44, 98-101.
- Otranto D, Dantas-Torres F, Breitschwerdt EB. Managing canine vector-borne diseases of zoonotic concern: part one. Trends Parasitol 2009, 25, 157-163. https://doi.org/10.1016/j.pt.2009.01.003
- Parola P, Roux V, Camicas JL, Baradji I, Brouqui P, Raoult D. Detection of ehrlichiae in African ticks by polymerase chain reaction. Trans R Soc Trop Med Hyg 2000, 94, 707-708. https://doi.org/10.1016/S0035-9203(00)90243-8
- Piratae S, Pimpjong K, Vaisusuk K, Chatan W. Molecular detection of Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli in stray dogs in Mahasarakham province, Thailand. Ann Parasitol 2015, 61, 183-187.
- Sabino CV, Weese JS. Contamination of multiple-dose vials in a veterinary hospital. Can Vet J 2006, 47, 779-782.
- Shaw SE, Day MJ, Birtles RJ, Breitschwerdt EB. Tickborne infectious diseases of dogs. Trends Parasitol 2001, 17, 74-80. https://doi.org/10.1016/S1471-4922(00)01856-0
- Suksawat J, Xuejie Y, Hancock SI, Hegarty BC, Nilkumhang P, Breitschwerdt EB. Serologic and molecular evidence of coinfection with multiple vector-borne pathogens in dogs from Thailand. J Vet Intern Med 2001, 15, 453-462. https://doi.org/10.1111/j.1939-1676.2001.tb01574.x
- Swofford DL. PAUP*. Phylogenetic analysis using parsimony (*and other methods). Version 4.0b10. Sinauer Associates, Sunderland, 2003.
- Taboada J, Lobetti R. Babesiosis. In: Green CE (ed.). Infectious Diseases of the Dog and Cat. 3rd ed. pp. 722-736, WB Saunders, Philadelpia, 2006.
- Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013, 30, 2725-2729. https://doi.org/10.1093/molbev/mst197
- Watanabe M, Oikawa T, Hiraoka H, Kaneko N, Itamoto K, Mizuno T, Okuda M, Inokuma H. Experimental inoculation of beagle dogs with Ehrlichia species detected from Ixodes ovatus. Vet Parasitol 2006, 136, 147-154. https://doi.org/10.1016/j.vetpar.2005.10.021
- Wong SSY, Teng JLL, Poon RWS, Choi GKY, Chan KH, Yeung ML, Hui JJY, Yuen KY. Comparative evaluation of a point-of-care immunochromatographic test SNAP 4Dx with molecular detection tests for vector-borne canine pathogens in Hong Kong. Vector Borne Zoonotic Dis 2011, 11, 1269-1277. https://doi.org/10.1089/vbz.2010.0265
- Ybanez AP. First molecular evidence of Ehrlichia canis infection in dogs with probable disease relapse in the Philippines. J Adv Vet Res 2014, 4, 184-188.
- Ybanez AP. First report on Anaplasma platys infection in a dog in the Philippines. Iran J Vet Med 2013, 7, 227-231.
- Ybanez AP, Perez ZO, Gabotero SR, Yandug RT, Kotaro M, Inokuma H. First molecular detection of Ehrlichia canis and Anaplasma platys in ticks from dogs in Cebu, Philippines. Ticks Tick Borne Dis 2012, 3, 288-293. https://doi.org/10.1016/j.ttbdis.2012.10.032
- Ybanez AP, Ybanez RHD, Talle MG, Liu M, Moumouni PFA, Xuan X. First report on Babesia vogeli infection in dogs in the Philippines. Parasitol Int 2017, 66, 813-815. https://doi.org/10.1016/j.parint.2016.10.001