doi: 10.14202/vetworld.2018.14-19
Share this article on [Facebook] [LinkedIn]
Article history: Received: 06-09-2017, Accepted: 05-12-2017, Published online: 12-01-2018
Corresponding author: Adrian P. Ybanez
E-mail: dr.adrianpybanez@gmail.com
Citation: Ybanez RHD, Ybanez AP, Arnado LLA, Belarmino LMP, Malingin KGF, Cabilete PBC, Amores ZRO, Talle MG, Liu M, Xuan X (2018) Detection of Ehrlichia, Anaplasma, and Babesia spp. in dogs in Cebu, Philippines, Veterinary World, 11(1): 14-19.Background: Ehrlichia, Anaplasma, and Babesia spp. are canine pathogens transmitted by the Rhipicephalus sanguineus tick which can cause varied clinical signs. These pathogens have been investigated in the Philippines, but coinfection has not been reported yet.
Aim: The aim of this study was to evaluate the presence of Ehrlichia/Anaplasma and Babesia spp. in Philippine dogs.
Materials and Methods: A total of 100 dogs from seven different veterinary establishments in Cebu, Philippines, were examined for Ehrlichia/Anaplasma and Babesia spp. infection using peripheral blood smear examination and polymerase chain reaction (PCR). Inclusion criteria included a history or presence of tick infestation, anemia, and/or thrombocytopenia. Clinical signs were recorded. Statistical analyses were performed between PCR positivity and clinical signs and hematological results.
Results: A total of 10 and 18 dogs were found to be positive for Ehrlichia/Anaplasma and Babesia spp., respectively. One animal was PCR positive for both pathogens, which is the first report of coinfection in the country. The most common clinical signs observed include inappetence (89%), lethargy (80%), thrombocytopenia (85%), and anemia (74%). Analyses revealed that inappetence (p=0.044) and weight loss (p=0.028) were found statistically significant with Ehrlichia/Anaplasma infection. Basophil (p=0.001) and eosinophil counts (p=0.000) were also found significantly different between Ehrlichia/Anaplasma spp.-positive and -negative dogs. On the other hand, differential monocyte count (p=0.009) was found significantly different between Babesia spp.-positive and -negative dogs.
Conclusion: The present study showed low infection rates of canine ehrlichiosis/anaplasmosis and babesiosis and provided additional evidence for the presence of the pathogens in the area.
Keywords: Babesia, Cebu, dogs, Ehrlichia/Anaplasma, Philippines.
1. Ybanez, A.P., Perez, Z.O., Gabotero, S.R., Yandug, R.T., Kotaro, M. and Inokuma, H. (2012) First molecular detection of Ehrlichiacanis and Anaplasmaplatys in ticks from dogs in Cebu, Philippines. Ticks Tickborne Dis., 3(5): 288-293.
2. Fourie, J.J., Stanneck, D., Luus, H.G., Beugnet, F., Wijnveld, M. and Jongejan, F. (2013) Transmission of Ehrlichiacanisby Rhipicephalussanguineus ticks feeding on dogs and on artificial membranes. Vet. Parasitol.,197(3): 595-603. [Crossref] [PubMed]
3. Ramos, R.A.N., Latrofa, M.S., Giannelli, A., Lacasella, V., Campbell, B.E., Dantas-Torres, F. and Otranto, D. (2014) Detection of Anaplasmaplatys in dogs and Rhipicephalussanguineus group ticks by a quantitative real-time PCR. Vet. Parasitol.,205(1): 285-288. [Crossref] [PubMed]
4. Ybanez, A.P., Ybanez, R.H.D., Yokoyama, N. and Inokuma, H. (2015) RNA polymerase sub-unit β (rpoB) characterization of Ehrlichiacanisdetected from dogs and Rhipicephalussanguineus ticks in Cebu, Philippines. Philippines. Vet. Arch., 85: 601-608.
5. Ybanez, A.P. and Inokuma, H. (2016) Anaplasma species of veterinary importance in Japan. Vet. World, 9(11): 1190. [Crossref] [PubMed] [PMC]
6. Dantas-Torres, F. (2010) Biology and ecology of the brown dog tick, Rhipicephalussanguineus. Parasit. Vectors, 3(1): 26. [Crossref] [PubMed] [PMC]
7. Al Izzi, S., Martin, D.S., Chan, R.Y. and Leutenegger, C.M. (2013) Babesiacanisvogeli, Ehrlichiacanis, and Anaplasma platys infection in a dog. Vet. Clin. Pathol.,42(4): 471-475. [Crossref] [PubMed]
8. Adamu, M., Troskie, M., Oshadu, D.O., Malatji, D.P., Penzhorn, B.L. and Matjila, P.T. (2014) Occurrence of tick-transmitted pathogens in dogs in Jos, Plateau State, Nigeria. Parasit. Vectors, 7(1): 119. [Crossref]
9. Costa, A.P.D., Costa, F.B., Labruna, M.B., Silveira, I., Moraes-Filho, J., Soares, J.F., Spolidorio, M.G. and Guerra, R.D.M. (2015) A serological and molecular survey of Babesiavogeli, Ehrlichiacanisand Rickettsia spp. among dogs in the state of Maranhao, Northeastern Brazil. Rev. Bras Parasitol. Vet., 24(1): 28-35. [Crossref] [PubMed]
10. Matjila, P.T., Leisewitz, A.L., Jongejan, F. and Penzhorn, B.L. (2008) Molecular detection of tick-borne protozoal and ehrlichial infections in domestic dogs in South Africa. Vet. Parasitol., 155(1): 152-157. [Crossref] [PubMed]
11. Kelly, P.J., Xu, C., Lucas, H., Loftis, A., Abete, J., Zeoli, F., Stevens, A., Jaegersen, K., Ackerson, K., Gessner, A. and Kaltenboeck B. (2013) Ehrlichiosis, babesiosis, anaplasmosis and hepatozoonosis in dogs from St. Kitts, West Indies. PLoS One, 8(1): e53450. [Crossref] [PubMed] [PMC]
12. Tanikawa, A., Labruna, M.B., Costa, A., Aguiar, D.M., Justiniano, S.V., Mendes, R.S., Melo, A.L.T., Alves, C.J. and Azevedo, S.S. (2013) Ehrlichiacanisin dogs in a semiarid region of Northeastern Brazil: Serology, molecular detection and associated factors. Res. Vet. Sci., 94(3): 474-477. [Crossref] [PubMed]
13. Ybanez, A.P., Ybanez, R.H.D., Villavelez, R.R., Malingin, H.P.F., Barrameda, D.N.M., Naquila, S.V. and Olimpos, S.M.B. (2016) Retrospective analyses of dogs found serologically positive for Ehrlichiacanisin Cebu, Philippines from 2003 to 2014. Vet. World, 9(1): 43-47. [Crossref] [PubMed] [PMC]
14. Cruz-Flores, M.J., Claveria, F.G., Verdida, R., Xuan, X. and Igarashi, I. (2008) First detection of Babesiagibsoni infection in Philippine stray dogs by immunochromatographic test (ICT). Vet. Arhiv., 78(2): 149.
15. Ybanez, A.P., Sivakumar, T., Ybanez, R.H.D., Ratilla, J.C., Perez, Z.O., Gabotero, S.R., Hakimi, H., Kawazu, S.I., Matsumoto, K., Yokoyama, N. and Inokuma, H. (2013) First molecular characterization of Anaplasmamarginale in cattle and Rhipicephalus(Boophilus) microplus ticks in Cebu, Philippines. J. Vet. Med. Sci., 75(1): 27-36. [Crossref]
16. Matsuu, A., Ono, S., Ikadai, H., Uchide, T., Imamura, S., Onuma, M. and Higuchi, S. (2005) Development of a SYBR green real-time polymerase chain reaction assay for quantitative detection of Babesiagibsoni (Asian genotype) DNA. J. Vet. Diagn. Invest., 17(6): 569-573. [Crossref] [PubMed]
17. Ybanez, A.P. (2014) First molecular evidence of Ehrlichiacanisinfection in dogs with probable disease relapse in the Philippines. J. Adv. Vet. Res., 4: 184-188.
18. Kordick, S.K., Breitschwerdt, E.B., Hegarty, B.C., Southwick, K.L., Colitz, C.M., Hancock, S.I., Bradley, J.M., Rumbough, R., Mcpherson, J.T. and MacCormack, J.N. (1999) Coinfection with multiple tick-borne pathogens in a walker hound kennel in North Carolina. J. Clin. Microbiol.,37(8): 2631-2638. [PubMed] [PMC]
19. Harrus, S. and Waner, T. (2011) Diagnosis of canine monocytotropicehrlichiosis (Ehrlichiacanis): An overview. Vet. J., 187(3): 292-296. [Crossref] [PubMed]
20. Akhtardanesh, B., Blourizadeh, H. and Ghanbarpour, R. (2010) Serological evidence of canine monocyticehrlichiosis in Iran. Comp. Clin. Pathol., 19: 469-474. [Crossref]
21. Buhles, W.C. Jr., Huxsoll, D.L. and Ristic, M. (1974) Tropical canine pancytopenia: Clinical, hematologic, and serologic response of dogs to Ehrlichiacanisinfection, tetracycline therapy, and challenge inoculation. J. Infect. Dis., 130(4): 357-367. [Crossref]
22. Elias, E. (1992) Diagnosis of ehrlichiosis from the presence of inclusion bodies or morulae of E can is. J. Small Anim. Pract., 33(11): 540-543. [Crossref]
23. McDade, J.E. (1990) Ehrlichiosis-a disease of animals and humans. J. Infect. Dis., 161: 609-617. [Crossref] [PubMed]
24. Dagnone, A.S., De Morais, H.S.A., Vidotto, M.C., Jojima, F.S. and Vidotto, O. (2003) Ehrlichiosis in anemic, thrombocytopenic, or tick-infested dogs from a hospital population in South Brazil. Vet. Parasitol., 117(4): 285-290. [Crossref]
25. Solano-Gallego, L., Trotta, M., Carli, E., Carcy, B., Caldin, M. and Furlanello, T. (2008) Babesiacanis and Babesiacanisvogeliclinicopathological findings and DNA detection by means of PCR-RFLP in blood from Italian dogs suspected of tick-borne disease. Vet. Parasitol.,157(3): 211-221. [Crossref] [PubMed]
26. Maia, M.G., Costa, R.T., Haddad, J.P., Passos, L.M. and Ribeiro, M.F. (2007) Epidemiological aspects of canine babesiosis in the semiarid area of the state of Minas Gerais, Brazil. Prev. Vet. Med., 79(2), 155-162. [Crossref] [PubMed]
27. Baticados, A.M., Baticados, W.N., Villarba, L.A., Carlos, E.T., Carlos, S. and Fajardo, P.V. (2011) PCR assay and microscopy for examination of mixed Ehrlichiacanis and Babesia spp. infection in bomb sniffing dogs and other canines in the national capital region, Philippines. Eur. J. Vet. Sci., 27: 111-115.
28. Corales, J.M.I., Viloria, V.V., Venturina, V.M. and Mingala, C.N. (2014) The prevalence of Ehrlichiacanis, Anaplasmaplatys and Babesia spp. in dogs in Nueva Ecija, Philippines based on multiplex polymerase chain reaction (mPCR) assay. Ann. Parasitol., 60: 267-272. [PubMed]
29. Das, M. and Konar, S. (2013) Clinical and hematological study of canine ehrlichiosis with other hemoprotozoan parasites in Kolkata, West Bengal, India. Asian Pac. J. Trop. Biomed., 3(11): 913-915. [Crossref]
30. Santos, F., Coppede, J.S., Pereira, A.L., Oliveira, L.P., Roberto, P.G., Benedetti, R.B. and Marins, M. (2009) Molecular evaluation of the incidence of Ehrlichiacanis, Anaplasmaplatys and Babesia spp. in dogs from RibeiraoPreto, Brazil. Vet. J., 179(1): 145-148. [Crossref] [PubMed]
31. Rojas, A., Rojas, D., Montenegro, V., Gutierrez, R., Yasur-Landau, D. and Baneth, G. (2014) Vector-borne pathogens in dogs from Costa Rica: First molecular description of Babesiavogeli and Hepatozooncanisinfections with a high prevalence of monocyticehrlichiosis and the manifestations of co-infection. Vet. Parasitol. 199(3): 121-128. [Crossref] [PubMed]
32. Foldvari, G., Hell, E. and Farkas, R. (2005) Babesiacanis in dogs from hungary: Detection by PCR and sequencing. Vet. Parasitol.,127(3): 221-226. [Crossref] [PubMed]
33. Duh, D., Tozon, N., Petrovec, M., StraSek, K. and AvSic-Zupanc, T. (2004) Canine babesiosis in Slovenia: Molecular evidence of Babesiacaniscanis and Babesiacanisvogeli. Vet. Res., 35(3): 363-368. [Crossref] [PubMed]
34. Huxsoll, D.L., Hildebrandt, P.K., Nims, R.M., Amyx, H.L. and Ferguson, J.A. (1970) Epizootiology of tropical canine pancytopenia. J. Wildlife Dis., 6(4): 220-225. [Crossref]
35. Waner, T. and Harrus, S. (2000) Canine monocyticehrlichiosis. In: Recent advances in canine infectious diseases. In: Carmichael, L.E., editor. International Veterinary Information Service. Available from: http://www.ivis.org/advances/infect_dis_carmichael/waner/ivis.pdf. Last accessed on 30-06-2017.
36. Ybanez, A.P., Ybanez, R.H.D., Talle, M.G., Liu, M., Moumouni, P.F.A. and Xuan, X. (2017) First report on Babesiavogeli infection in dogs in the Philippines. Parasitol. Int., 66(1): 813-815. [Crossref] [PubMed]
37. Nakaghi, A.C.H., Machado, R.Z., Costa, M.T., Andre, M.R. and Baldani, C.D. (2008) Canine ehrlichiosis: Clinical, hematological, serological and molecular aspects. Ciencia Rural., 38: 766-770. [Crossref]
38. Waner, T., Harrus, S., Bark, H., Bogin, E., Avidar, Y. and Keysary, A. (1997) Characterization of the subclinical phase of canine ehrlichiosis in experimentally infected beagle dogs. Vet. Parasitol., 69: 307-317. [Crossref]
39. Harrus, S., Waner, T., Keysary, A., Aroch, I., Voet, H. and Bark, H. (1998) Investigation of splenic functions in canine monocyticehrlichiosis. Vet. Immunol. Immunopath., 62: 15-27. [Crossref]
40. Bulla, C., Takahira, R.K., Araujo J.P. Jr., Aparecida, T.L., Lopes, R.S. and Wiedmeyer, C.E. (2004) The relationship between the degree of thrombocytopenia and infection with Ehrlichiacanisin an endemic area. Vet. Res., 35(1): 141-146. [Crossref] [PubMed]
41. Harrus, S., Waner, T., Aizenberg, I. and Bark, H. (1998) Therapeutic effect of doxycycline in experimental subclinical canine monocyticehrlichiosis: Evaluation of a 6-week course. J. Clin. Microbiol., 36: 2140-2142. [PubMed] [PMC]
42. Rene-Martellet, M., Chene, J., Chabanne, L., Chalvet-Monfray, K. and Bourdoiseau, G. (2013) Clinical signs, seasonal occurrence and causative agents of canine babesiosis in France: Results of a multiregional study. Vet. Parasitol.,197(1): 50-58. [Crossref] [PubMed]
43. Salem, N.Y. and Farag, H.S. (2014) Clinical, hematologic, and molecular findings in naturally occurring Babesiacanisvogeli in Egyptian dogs. Vet. Med. Int., 2014: 270345. [Crossref] [PubMed] [PMC]
44. Shaw, S.E., Day, M.J., Birtles, R.J. and Breitschwerdt, E.B. (2001) Tick-borne infectious diseases of dogs. Trends. Parasitol., 17(2): 74-80. [Crossref]
45. Lasta, C.S., Santos, A.P.D., Messick, J.B., Oliveira, S.T., Biondo, A.W., Vieira, R.F.D., Dalmolin, M.L. and Gonzalez, F.H.D. (2013) Molecular detection of Ehrlichiacanis and Anaplasmaplatys in dogs in Southern Brazil. Rev. Bras. Parasitol. Vet.,22(3): 360-366. [Crossref] [PubMed]
46. Zygner, W., Gojska, O., Rapacka, G., Jaros, D. and Wedrychowicz, H. (2007) Hematological changes during the course of canine babesiosis caused by large Babesia in domestic dogs in Warsaw (Poland). Vet. Parasitol.,145(1): 146-151. [Crossref] [PubMed]
47. Latrofa, M.S., Dantas-Torres, F., Giannelli, A. and Otranto, D. (2014) Molecular detection of tick-borne pathogens in Rhipicephalussanguineus group ticks. Ticks Tickborne Dis., 5(6): 943-946. [Crossref] [PubMed]
48. Cicuttin, G.L., Brambati, D.F., Eugui, J.I.R., Lebrero, C.G., De Salvo, M.N., Beltran, F.J., Dohmen, F.E.G., Jado, I. and Anda, P. (2014) Molecular characterization of Rickettsia massiliae and Anaplasma platys infecting Rhipicephalussanguineus ticks and domestic dogs, Buenos Aires (Argentina). Ticks Tick-borne Dis., 5(5): 484-488. [Crossref] [PubMed]
49. Vargas-Hernandez, G., Andre, M.R., Faria, J.L.M., Munhoz, T.D., Hernandez-Rodriguez, M., Machado, R.Z. and Tinucci-Costa, M. (2012) Molecular and serological detection of Ehrlichiacanisand Babesiavogeli in dogs in Colombia. Vet. Parasitol.,186(3): 254-260. [Crossref] [PubMed]
50. Rani, P.A.M.A., Irwin, P.J., Coleman, G.T., Gatne, M. and Traub, R.J. (2011) A survey of canine tick-borne diseases in India. Parasites Vectors, 4(1): 141. [Crossref] [PubMed] [PMC]
51. Hii, S.F., Kopp, S.R., Thompson, M.F., O'leary, C.A., Rees, R.L. and Traub, R.J. (2012) Canine vector-borne disease pathogens in dogs from south-east Queensland and north-east Northern Territory. Aust. Vet. J., 90(4): 130-135. [Crossref] [PubMed]
52. Gal, A., Harrus, S., Arcoh, I., Lavy, E., Aizenberg, I., Mekuzas-Yisaschar, Y. and Baneth, G. (2007) Coinfection with multiple tick-borne and intestinal parasites in a 6-week-old dog. Can. Vet. J., 48: 619-622. [PubMed] [PMC]
53. de Caprariisa, D., Dantas-Torresa, F., Capellib, G., Menckec, N., Stanneckc, D., Breitschwerdt, E.B. and Otrantoa, D. (2011) Evolution of clinical, haematological and biochemical findings in young dogs naturally infected by vector-borne pathogens. Vet. Microbiol., 149: 206-212. [Crossref] [PubMed]