SHORT COMMUNICATION

 

Confirmation of occurrence of Babesia vogeli in a dog in Windhoek, central Namibia

 

 

Barend L. Penzhorn^I; Ilse Vorster^I; Gernot Redecker^II; Marinda C. Oosthuizen^I

^IDepartment of Veterinary Tropical Diseases, University of Pretoria, South Africa
^IIWindhoek Animal Hospital, Windhoek, Namibia

Correspondence

 

 

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ABSTRACT

Although there is evidence of high seroprevalence of antibodies to Babesia spp. in dogs in central Namibia, clinical babesiosis is rarely diagnosed. Rhipicephalus sanguineus sensu lato, the vector of Babesia vogeli, is common in Namibia while Haemaphysalis elliptica, the vector of the highly virulent but morphologically indistinguishable Babesia rossi, has rarely been recorded, mainly in northern Namibia. On the basis of vector occurrence, clinical cases of canine babesiosis in Windhoek, central Namibia, have been ascribed to B. vogeli. DNA extracted from a blood smear made from a sick dog was subjected to the reverse line blot hybridisation assay. The polymerase chain reaction amplicons hybridised with the B. vogeli-specific probe, but not with the Babesia canis- and B. rossi-specific probes. Although attempts at cloning and sequencing of the full-length 18S rRNA gene were unsuccessful, we can confirm that B. vogeli occurs in central Namibia.

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Introduction

Canine babesiosis is caused by either 'large' or 'small' piroplasms. The latter, primarily Babesia gibsoni, are not of major concern in southern Africa and are not further considered here (Matjila et al. 2007). Differences in clinical manifestation, immunology and vector specificity of isolates of 'large' babesias from various geographic regions were reported at an early stage (e.g. Brumpt 1919; Christophers 1907; Laveran & Nattan-Larrier 1913; Robertson 1901), but were mostly overlooked, and the organism was generally referred to as Babesia canis sensu lato. The situation changed when three groups of B. canis s. l. were distinguished and a new nomenclature was proposed (Uilenberg et al. 1989). The 'large' piroplasms are a complex of at least three morphologically similar but genetically distinct vector-specific species: B. canis, Babesia rossi and Babesia vogeli (Zahler et al. 1998). The pre-1989 literature on canine babesiosis should therefore be critically evaluated to determine which specific taxon is involved.

Babesia rossi, transmitted by Haemaphysalis elliptica and presumably also by Haemaphysalis leachi (Apanaskevich, Horak & Camicas 2007), is restricted to sub-Saharan Africa and causes the most virulent manifestation of canine babesiosis. The less virulent B. canis sensu stricto, transmitted by Dermacentor reticulatus, is restricted to Europe (Brumpt 1919). The least virulent species, B. vogeli, is transmitted by Rhipicephalus sanguineus sensu lato, which has a global distribution, but is most prevalent in tropical and subtropical regions (Gray et al. 2013).

Canine babesiosis is regarded as a major problem in South Africa, where it has been well studied (Jacobson 2006; Schoeman 2009). The vast majority of cases are caused by B. rossi: at the Onderstepoort Veterinary Academic Hospital (OVAH), only 5 of 350 cases (1.4%) were attributed to B. vogeli, the remaining 98.6% of the patients being positive for B. rossi (Matjila et al. 2008). At the OVAH, H. elliptica (reported as H. leachi) was collected from 304 of 395 dogs (77.0%) diagnosed with canine babesiosis, while R. sanguineus was collected from 146 (37%) of these dogs (Horak 1995).

Initial reports indicated that canine babesiosis is not common in Namibia (Schneider 1994). In a more recent study, Stübe (2004) identified Babesiapiroplasms in 27 of 600 blood smears (4.5%) made from dogs in the Okahandja District of central Namibia. Using an enzyme-linked immunosorbent assay (ELISA) with B. canis antigen, 69% of the 600 dogs were found to be seropositive (Stübe 2004). This test is known to cross-react with antibodies to B. vogeli, while the IFA test using this antigen cross-reacts with both B. rossi and B. vogeli (Dyachenko et al. 2012; Jongejan et al. 2011; Pantchev et al. 2015). This would suggest either (1) an endemically stable situation, such as typically occurring with bovine babesiosis, or (2) the presence of a parasite of relatively low virulence.

Although we found no published reference to the occurrence of ticks on dogs in Windhoek, R. sanguineus s. l. was the most numerous tick species infesting dogs in Okahandja, Gobabis, Mariental and Rosh Pinah, Namibia (Matthee et al. 2010; Stübe 2004). All 796 ticks collected from dogs at Okahandja were R. sanguineus (Stübe 2004); 99.2% of the ticks collected at the other three localities were also of this species (Matthee et al. 2010). Among 899 ticks collected from dogs, Matthee et al. (2010) identified a single male of the genus Haemaphysalis; the species could not be determined. Haemaphysalis elliptica (reported as H. leachi), usually associated with more mesic conditions, has been reported from northern Namibia and once from Karasburg, southern Namibia (Walker 1991).

On the basis of the abundance of R. sanguineus s. l., Noden and Soni (2015) ascribed clinical canine babesiosis cases in Namibia to B. vogeli. We can confirm that B. vogeli occurs in Namibia.

In February 2015, a dog presented at Windhoek Animal Hospital showed typical clinical signs of babesiosis. A blood smear was made, and large babesias were seen (Figure 1). The dog received the usual treatment and made an uneventful recovery. A stained blood smear was sent to the Department of Veterinary Tropical Diseases (DVTD) for confirmation of the diagnosis.

 

 

At the DVTD, the dried blood was removed from the slide. DNA was extracted using the QIAamp^® DNA Mini Kit (QIAGEN, Whitehead Scientific, South Africa) and subjected to the reverse line blot (RLB) hybridisation assay. A 460 bp - 520 bp fragment of the V4 variable region of the parasite 18S rRNA gene was amplified by polymerase chain reaction (PCR) using the Theileria and Babesia genus-specific primers RLB F2 [5'-GAC ACA GGG AGG TAG TGA CAA G-3'] and biotin-labelled RLB R2 [5'-Biotin-CTA AGA ATT TCA CCT CTA ACA GT-3'] (Nijhof et al. 2003, 2005). The PCR products were analysed using the RLB hybridisation assay, first described by Gubbels et al. (1999). The RLB membrane contained Theileria and Babesia genus- and species-specific probes, including B. canis (5'-TGC GTT GAC GGT TTG AC-3'), B. rossi (5'-CGG TTT GTT GCC TTT GTG-3') and B. vogeli (5'-AGC GTG TTC GAG TTT GCC-3') (Matjila et al. 2004).

On RLB, the PCR amplicons hybridised only with the B. vogeli-specific probe. Although attempts at cloning and sequencing of the full-length 18S rRNA gene were unsuccessful, we can confirm that B. vogeli occurs in central Namibia. This does not rule out possible occurrence of B. rossi, especially in the more mesic north-eastern parts of Namibia, including the Zambezi (previously Caprivi) region, where H. elliptica could occur since it is found in the adjoining Okavango region of Botswana (Walker 1991).

In sub-Saharan Africa, B. vogeli was first reported from clinically normal dogs in South Africa (Matjila et al. 2004). Its presence had probably been overlooked because of the preponderance of infections of the highly virulent B. rossi (Matjila et al. 2008). Babesia vogeli has also been reported from Sudan (Oyamada et al. 2005), Nigeria (Adamu et al. 2014; Kamani et al. 2013; Sasaki et al. 2007) and Cape Verde (Götsch et al. 2009). In Zimbabwe, B. vogeli has been reported from captive lions (Panthera leo), wild cats (Felis lybica) and servals (Leptailurus serval) and could presumably also occur in dogs (Kelly et al. 2014).

Rhipicephalus sanguineus s. l., the only known vector of B. vogeli, is now thought to be a species-complex comprising at least 17 sibling species, which may differ in vector capacity (Dantas-Torres et al. 2013; Dantas-Torres & Otranto 2015). Domestication of the dog benefitted this tick, which probably evolved as a parasite of burrowing carnivores in warm climates (Gray et al. 2013). Because of the worldwide spread of humans and dogs, this tick now has a global distribution. Rhipicephalus sanguineus s. l. is incriminated as vector of Hepatozoon canis and Ehrlichia canis in addition to B. vogeli. These pathogens could therefore be expected to occur throughout the geographic distribution of the vector (Gray et al. 2013). Indeed, 53.8% of 106 dogs from Windhoek, Namibia, were seropositive on a solid-phase dot ELISA based on a crude antigen of E. canis (Israeli strain) (Manyarara et al. 2015).

 

Acknowledgements

We thank Mr Louis Taljaard for the photomicrograph and Prof. Georg von Sampson-Himmelstjerna of the Free University, Berlin, for furnishing a copy of Julia Stübe's Dr Med Vet dissertation.

Competing interests

The authors declare that they have no financial or personal relationships that may have inappropriately influenced them in writing this article.

Authors' contributions

B.L.P. obtained the specimen and wrote the report. I.V. performed the DNA extraction and RLB. M.C.O. advised on interpretation of results, and contributed to the manuscript. G.R. was the clinician handling the case.

 

References

Adamu, M., Troskie, M., Oshadu, D.O., Malatji, D.P., Penzhorn, B.L. & Matjila, P.T., 2014, 'Occurrence of tick-transmitted pathogens in dogs in Jos, Plateau State, Nigeria', Parasites & Vectors 7, Art. #119. http://dx.doi.org/10.1186/1756-3305-7-119        [ Links ]

Apanaskevich, D.A., Horak, I.G. & Camicas, J.-L., 2007, 'Redescription of Haemaphysalis (Rhipistoma) elliptica (Koch, 1844), an old taxon of the Haemaphysalis (Rhipistoma) leachi group from East and southern Africa, and of Haemaphysalis (Rhipistoma) leachi (Audouin, 1826) (Ixodida, Ixodidae)', Onderstepoort Journal of Veterinary Research 74, 181-208. http://dx.doi.org/10.4102/ojvr.v74i3.122        [ Links ]

Brumpt, E., 1919, 'Transmission de la piroplasmose canine française par le Dermacentor reticulatus. Embolies parasitaires dans les capillaires de l'encéphale', Bulletin de la Société de Pathologie Exotique 12, 651-664.         [ Links ]

Christophers, S.R., 1907, 'Preliminary note on the development of Piroplasma canis in the tick', British Medical Journal 1(2402), 76-78. http://dx.doi.org/10.1136/bmj.1.2402.76        [ Links ]

Dantas-Torres, F., Latrofa, M.S., Annoscia, G., Giannelli, A., Parisi, A. & Otranto, D., 2013, 'Morphological and genetic diversity of Rhipicephalus sanguineus sensu lato from the New and Old Worlds', Parasites & Vectors 6, Art. #213. http://dx.doi.org/10.1186/1756-3305-6-213        [ Links ]

Dantas-Torres, F. & Otranto, D., 2015, 'Further thoughts on the taxonomy and vector role of Rhipicephalus sanguineus group ticks', Veterinary Parasitology 208, 9-13. http://dx.doi.org/10.1016/j.vetpar.2014.12.014        [ Links ]

Dyachenko, V., Pantchev, N., Balzer, H.-J., Meyersen, A. & Straubinger, R., 2012, 'First case of Anaplasma platys infection in a dog in Croatia', Parasites & Vectors 5, 49. http://dx.doi.org/10.1186/1756-3305-5-49        [ Links ]

Götsch, S., Leschnik, M., Duscher, G., Burgstaller, J.P., Wille-Piazzai, W. & Joachim, A. 2009, 'Ticks and haemoparasites of dogs from Praia, Cape Verde', Veterinary Parasitology 166, 171-174. http://dx.doi.org/10.1016/j.vetpar.2009.08.009        [ Links ]

Gray, J., Dantas-Torres, F., Astrada-Peña, A. & Levin, M., 2013, 'Systematics and ecology of the brown dog tick, Rhipicephalus sanguineus', Ticks and Tick-Borne Diseases 4, 171-180 http://dx.doi.org/10/1016/j.ttbdis.2012.12.003        [ Links ]

Gubbels, J.M., De Vos, A.P., Van Der Weide, M., Viseras, J., Schouls, L.M., De Vries, E. et al., 1999, 'Simultaneous detection of bovine Theileria and Babesia species by reverse line blot hybridization', Journal of Clinical Microbiology 37, 1782-1789.         [ Links ]

Horak, I.G., 1995, 'Ixodid ticks collected at the Faculty of Veterinary Science, Onderstepoort, from dogs diagnosed with Babesia canis infection', Journal of the South African Veterinary Association 66, 170-171.         [ Links ]

Jacobson, L., 2006, 'The South African form of severe and complicated canine babesiosis: Clinical advances 1994-2004', Veterinary Parasitology 138, 126-139, http://dx.doi.org/10.1016/j.vetpar.2006.01.047        [ Links ]

Jongejan, F., Fourie, J.J., Chester, T., Manavella, C., Mallouk, Y., Pollmeier, M.G. et al., 2011, 'The prevention of transmission of Babesia canis canis by Dermacentor reticulatus ticks to dogs using a novel combination of fipronil, amitraz and (S)-methoprene', Veterinary Parasitology 179, 343-350 http://dx.doi.org/10.1016/j.vetpar.2011.03.047        [ Links ]

Kamani, J., Baneth, G., Mumcuoglu, K.Y., Waziri, N.E., Eyal, O., Guthmann, Y. et al., 2013, 'Molecular detection and characterization of tick-borne pathogens in dogs and ticks from Nigeria', PLoS Neglected Tropical Diseases 7(3), Art. #e2108. http://dx.doi.org/10.1371/journal.pntd.0002108        [ Links ]

Kelly, P., Marabini, L., Dutlow, K., Zhang, J.L., Loftis, A. & Wang, C.M., 2014, 'Molecular detection of tick-borne pathogens in captive wild felids, Zimbabwe', Parasites & Vectors 7, Art. #UNSP 514. http://dx.doi.org/10.1186/s13071-014-0514-6        [ Links ]

Laveran, A. & Nattan-Larrier, 1913, 'Piroplasmoses canines d'Europe et d'Afrique', Annales de l'Institut Pasteur 27, 701-717.         [ Links ]

Manyarara, R., Tubbesing, U., Soni, M. & Noden, B.H., 2015, 'Serodetection of Ehrlichia canis amongst dogs in central Namibia', Journal of the South African Veterinary Association 86(1), Art. # 1272, 1-3. http://dx.doi.org/10.4102/jsava.v86i1.1272        [ Links ]

Matjila, P.T., Leisewitz, A.L., Jongejan, F. & Penzhorn, B.L., 2008, 'Molecular detection of tick-borne protozoal and ehrlichial infections in domestic dogs in South Africa', Veterinary Parasitology 155, 152-157. http://dx.doi.org/10.1016/j.vetpar.2008.04.012        [ Links ]

Matjila, P.T., Penzhorn, B.L., Bekker, C.P.J., Nijhof, A.M. & Jongejan, F., 2004, 'Confirmation of occurrence of Babesia canis vogeli in domestic dogs in South Africa', Veterinary Parasitology 122, 119-125. http://dx.doi.org/10.1016/j.vetpar.2004.03.019        [ Links ]

Matjila, P.T., Penzhorn, B.L., Leisewitz, A.L., Bhoora, R. & Barker, R., 2007, 'Molecular characterization of Babesia gibsoni infection from a Pit-bull terrier pup recently imported into South Africa', Journal of the South African Veterinary Association 78, 2-5, http://dx.doi.org/10.4102/jsava.v78i1.277        [ Links ]

Matthee, S., Lovely, C., Gaugler, A., Beeker, R., Venter, H.R. & Horak, I.G., 2010, 'Ixodid ticks on domestic dogs in the Northern Cape Province of South Africa and in Namibia', Journal of the South African Veterinary Association 81, 126-128. http://dx.doi.org/10.4102/jsava.v81i2.125        [ Links ]

Nijhof, A., Penzhorn, B.L., Lynen, G., Mollel, J.O., Bekker, C. & Jongejan, F., 2003, 'Babesia bicornis sp. n. and Theileria bicornis sp. n.: Tick-borne parasites associated with mortality in the black rhinoceros (Diceros bicornis)', Journal of Clinical Microbiology 41, 2249-2254. http://dx.doi.org/10.1128/JCM.41.5.2249-2254.2003        [ Links ]

Nijhof, A.M., Pillay, V., Steyl, J., Prozesky, L., Stoltsz, W.H., Lawrence, J.A. et al., 2005, 'Molecular characterization of Theileria species associated with mortality in four species of African antelopes', Journal of Clinical Microbiology 43, 5907-5911. http://dx.doi.org/10.1128/JCM.43.12.5907-5911.2005        [ Links ]

Noden, B.H. & Soni, M., 2015, 'Vector-borne diseases of small companion animals in Namibia: Literature review, knowledge gaps and opportunity for a one health approach', Journal of the South African Veterinary Association 86(1), Art. #1307, 1-7. http://dx.doi.org/10.4102/jsava/v86i1.1307        [ Links ]

Oyamada, M., Davoust, B., Boni, M., Dereure, J., Bucheton, B., Hammad, A. et al., 2005, 'Detection of Babesia canis rossi, B. canis vogeli, and Hepatozoon canis in dogs in a village of eastern Sudan by using a screening PCR and sequencing methodologies', Clinical and Diagnostic Laboratory Immunology 12, 1343-1346. http://dx.doi.org/10.1128/CDLI.12.11.1343-1346.2005        [ Links ]

Pantchev, N., Pluta, S., Huisinga, E., Nather, S., Scheufelen, M., Vrhovec, M.G. et al., 2015, 'Tick-borne diseases (borreliosis, anaplasmosis, babesiosis) in German and Austrian dogs: Status quo and review of distribution, transmission, clinical findings, diagnostics and prophylaxis', Parasitology Research 114 (Suppl 1), S19-S54. http://dx.doi.org/10.10007/s00436-015-4513-0        [ Links ]

Robertson, W., 1901, 'Malignant jaundice in the dog', Journal of Comparative Pathology and Therapeutics 14, 327-336. http://dx.doi.org/10.1016/S0368-1742(01)80065-5        [ Links ]

Sasaki, M., Omobowale, O., Tozuka, M., Ohta, K., Matsuu, A., Nottidge, H.O. et al., 2007, 'Molecular survey of Babesia canis in dogs in Nigeria', Journal of Veterinary Medical Science 69, 1191-1193. http://dx.doi.org/10.1292/jvms.69.1191        [ Links ]

Schneider, H.P., 1994, Animal health and veterinary medicine in Namibia, Agrivet, Windhoek.         [ Links ]

Schoeman, J.P., 2009, 'Canine babesiosis', Onderstepoort Journal of Veterinary Research 76, 59-66. http://dx.doi.org/10.4102/ojvr.v76i1.66        [ Links ]

Stübe, J., 2004, Investigations on the prevalence of ehrlichiosis and babesiosis in dogs in Namibia with special consideration of the housing conditions, Hochschulschrift, Freie Universität Berlin, Berlin.         [ Links ]

Uilenberg, G., Franssen, F.F., Perie, N.M. & Spanjer, A.A., 1989, 'Three groups of Babesia canis distinguished and a proposal for nomenclature', Veterinary Quarterly 11, 33-40. http://dx.doi.org/10.1080/01652176.1989.9694194        [ Links ]

Walker, J.B., 1991, 'A review of the ixodid ticks (Acari, Ixodidae) occurring in Southern Africa', Onderstepoort Journal of Veterinary Research 58, 81-105.         [ Links ]

Zahler, M., Schein, E., Rinder, H. & Gothe, R., 1998, 'Characteristic genotypes discriminate between Babesis canis isolates of differing vector specificity and pathogenicity to dogs', Parasitology Research 84, 544-548. http://dx.doi.org/10.1007/s004360050445        [ Links ]

 

 

Correspondence:
Barend Penzhorn
banie.penzhorn@up.ac.za

Received: 29 Apr. 2016
Accepted: 18 Sept. 2016
Published: 26 Oct. 2016