Skip to main content

Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland



Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland.

Case presentation

A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h.


This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.


Tick-borne diseases (TBD) constitute a diversified group of diseases of increasing importance in human and veterinary medicine [1]. As showed by the observation of many authors, in Europe ticks are considered the most important of the arthropod zoonotic vectors [2,3,4], that are able to transmit such pathogens as Anaplasma spp., Babesia/Theileria spp. or Borrelia spp. Anaplasma spp. are pathogenic for several animal host species [5], while Anaplasma phagocytophilum is an emerging human pathogen in the USA and Europe [6, 7]. The clinical form of the disease is rarely reported in wild animals in captivity [8,9,10].

The aim of this study was to present the clinical case of granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland.

Case presentation

The observation took place in March 2020. The animal concerned was a female ring-tailed lemur (Lemur catta), 4 years old, with a body weight of 4.2 kg, with signs of anorexia, weakness, epistaxis and uncoordinated gait. These clinical signs appeared four days before the animal was brought to the clinic. During a clinical examination two adult female Ixodes ricinus ticks were removed from the animal’s body. The ticks were identified on the basis of morphology using taxonomic keys [11]. The lemur came from a zoo in eastern Poland. The animal lived in a group of 6 lemurs. Two months before she had received fenbendazole (50 mg/kg p.o. for 3 days) as deworming treatment, but no ectoparasite prophylaxis had been applied. The animal was clinically examined and blood samples were collected for biochemical, haematological and molecular tests for tick-borne diseases (babesiosis/theileriosis/anaplasmosis/ehrlichiosis). The ticks found on the animal’s body were also tested for the above diseases using molecular methods.

DNA extractions from the blood samples and ticks for molecular tests were performed using a commercial DNA Genomic kit (A&A Biotechnology Gdańsk, Poland) following the manufacturer’s instructions. Subsequent PCR tests were performed according to the methods described by Skotarczak et al. [12], Altay et al. [13] and Adaszek and Winiarczyk [14] (Table 1). The final identification of tick-borne pathogens was performed by sequencing PCR products.

Table 1 PCR conditions and primers used in the PCR protocols for detecting Anaplasma/Ehrlichia spp., Borrelia burgdorferi sensu lato and Babesia/Theileria spp

The haematological and biochemical test results did not reveal any abnormalities, except for thrombocytopaenia (PLT = 61 × 109/l; range 165–685 × 109/l) [15]. The microscopic examination of the stained blood smear (Giemsa method) revealed morulae in the cytoplasm of circulating neutrophils suggestive of acute granulocytic anaplasmosis (Fig. 1). The PCR test revealed Anaplasma DNA in the lemur’s blood and in I. ricinus ticks collected from the lemur’s body (Fig. 2). The analysis of PCR product sequencing identified the Rickettsia species as A. phagocytuphilum GU183908 (100 % homology). Based on the microscopic blood smears and the molecular test results, the disease was caused by Anaplasma phagocytophilum infection. The treatment started with doxycycline (2.5 mg/kg p.o., b.i.d.) administered for three weeks. 24 h after the initiation of the treatment the lemur’s condition improved significantly: her appetite increased and normal gait was restored. Three days later the epistaxis had subsided. Two weeks following the initiation of the antibiotic treatment a sample of the animal’s blood was collected for a quick test to detect the presence of A. phagocytophilum antibodies (VetExpert, CaniV-4 Poland). The test result was positive. A control PCR test carried out after the next three weeks (according to the same procedure as previous) did not reveal genetic material of A. phagocytophilum in the animal’s blood. Also, no DNA of bacteria was found in the blood of the other five lemurs from the same institution. Six months after the beginning of therapy the antibodies for A. phagocytophilum were still be present in the animal’s blood.

Fig. 1

Presence of morulae of A. phagocytophilum inside neutrophil cell (marked with an arrow)

Fig. 2

PCR amplification of a partial sequence of A. phagocytophilum 16 S RNA gene (product size 247 bp). Legend: lane 1 - molecular weight marker = 100 bp; lane 2 - negative control; lane 3 - positive control (A. phagocytophilum from human blood – National Reference Center for Borreliae of Max von Pettenkofer Institute of Ludwig Maximilian University Munich); lane 4 – amplification product of blood sample of lemur; lanes 5–7 - amplification products of ticks samples

Discussion and conclusions

This article presents the first clinical case of granulocytic anaplasmosis in a lemur in Europe. A. phagocytophilum is one of the most prevalent tick-transmitted animal and human pathogen [7]. The main clinical disorders observed in the course of granulocytic anaplasmosis are fever, thrombocytopenia and lameness. Ticks and wildlife are the main reservoirs of these bacteria [8], but clinical disease in free-ranging as well as in captive wild animals appears to be rare. However wildlife may play a role in the transmission and maintenance of granulocytic anaplasmosis, either acting as a reservoir of the bacteria or amplifying host for human or domestic animals [16]. Therefore, it is important to identify the potential hosts and characterise the role in the epidemiology of various animal species in this disease in order to adequately evaluate the potential risks and to design proper strategies of control.

The main vector of the microorganisms in Europe is the tick Ixodes ricinus [17]. There are only a few specific reports regarding A. phagocytophilum infection in lemurs. Specific antibodies against A. phagocytophilum were found in the serum of lemurs from St. Catherine’s Island, Georgia, USA [18], whereas screening tests for infections, including A. phagocytophilum, conducted in the lemur population of Madagascar did not confirm a single case of the disease [19]. In Poland the disease was previously detected in horses [14], dogs [20] and cats [21], but never in exotic animals.

The definitive diagnosis of A. phagocytophilum infection in a ring-tailed lemur was confirmed by results of PCR and sequencing. The 16 S rRNA gene fragment of bacteria detected in the blood of the patient, as well as in the tick organism collected from the lemur’s body showed 100 % similarity with GU183908 uncultured Anaplasma species clone Lublin-1 from previous studies [14]. This suggests an endemic occurrence of this microorganism strain in Poland. The description of the presented case indicates that A. phagocytophilum infection must also be considered in differential diagnosis in exotic animals living in Poland, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

Availability of data and materials

All data generated or analysed during this study are included in this published article.



Tick-borne diseases




Milligrams per kilogram

p.o. :

Per os


Deoxyribonucleic Acid


Polymerase Chain Reaction

s.l. :

Sensu lato

spp. :

Species pluralis




Bis in die


Ribosomal Ribonucleic Acid


  1. 1.

    Vayssier-Taussat M, Kazimirova M, Hubalek Z, Hornok S, Farkas R, Cosson JF, et al. Emerging horizons for tick-borne pathogens: from the ‘one pathogen - one disease’ vision to the pathobiome paradigm. Future Microbiol. 2015;10:2033–43.

    CAS  Article  Google Scholar 

  2. 2.

    Munderloh U. Comparative studies in tick-borne diseases in animals and humans. Vet Sci. 2017;4:E32.

    Article  Google Scholar 

  3. 3.

    Andersson MO, Marga G, Banu T, Dobler G, Chitimia-Dobler L. Tick-borne pathogens in tick species infesting humans in Sibiu County, central Romania. Parasitol Res. 2018;117:1591–7.

    Article  Google Scholar 

  4. 4.

    Krämer F, Hüsken R, Krüdewagen EM, Deuster K, Blagburn B, Straubinger RK, et al. Prevention of transmission of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum by Ixodes spp. ticks in dogs treated with the Seresto® collar (imidacloprid 10 % + flumethrin 4.5 %). Parasitol Res. 2020;119:299–315.

  5. 5.

    Battilani M, De Arcangeli S, Balboni A, Dondi F. Genetic diversity and molecular epidemiology of Anaplasma. Infect Genet Evol. 2017;49:195–211.

    CAS  Article  Google Scholar 

  6. 6.

    Doudier B, Olano J, Parola P, Brouqui P. Factors contributing to the emergence of Ehrlichia and Anaplasma spp. as human pathogens. Vet Parasitol. 2010;167:149–54.

    CAS  Article  Google Scholar 

  7. 7.

    Kocan KM, de la Fuente J, Coburn LA. Insights into the development of Ixodes scapularis: a resource for research on a medically important tick species. Parasit Vectors. 2015;8:592.

    Article  Google Scholar 

  8. 8.

    Nelder MP, Reeves WK, Adler PH, Wozniak A, Wills W. Ectoparasites and associated pathogens of free-roaming and captive animals in zoos in South Carolina. Vector Borne Zoonotic Dis. 2009;9:469–77.

    Article  Google Scholar 

  9. 9.

    Leschnik M, Kirtz G, Virányi Z, Wille-Piazzai W, Duscher G. Acute granulocytic anaplasmosis in a captive timber wolf (Canis lupus occidentalis). J Zoo Wildl Med. 2012;43:645–8.

    Article  Google Scholar 

  10. 10.

    Sim RR, Joyner PH, Padilla LR, Anikis P, Aitken-Palmer C. Clinical disease associated with Anaplasma phagocytophilum infection in captive Przewalski’s horses (Equus ferus Przewalskii). J Zoo Wildl Med. 2017;48:497–505.

    Article  Google Scholar 

  11. 11.

    Estrada-Peña A, Bouattour A, Camicas JL, Walker AR. Tick of domestic animals in Mediterranean region. A guide to identification of species. Zagaroza: University of Zaragoza; 2004.

    Google Scholar 

  12. 12.

    Skotarczak B, Wodecka B, Rymaszewska A, Sawczuk M, Maciejewska A, Adamska M, et al. Prevalence of DNA and antibodies to Borrelia burgdorferi sensu lato in dogs suspected of borreliosis. Ann Agric Environ Med. 2005;12:199–205.

    CAS  PubMed  Google Scholar 

  13. 13.

    Altay K, Aydin MF, Dumanli N, Aktas M. Molecular detection of Theileria and Babesia infections in cattle. Vet Parasitol. 2008;158:295–301.

    CAS  Article  Google Scholar 

  14. 14.

    Adaszek Ł, Winiarczyk S. Identification of Anaplasma spp. rickettsia isolated from horses from clinical disease cases in Poland. Zoonoses Public Health. 2011;58:514–8.

    CAS  Article  Google Scholar 

  15. 15.

    Williams CV, Van Steenhouse JL, Bradley JM, Hancock SI, Hegarty BC, Breitschwerdt EB. Naturally occurring Ehrlichia chaffeensis infection in two prosimian primate species: ring-tailed lemurs (Lemur catta) and ruffed lemurs (Varecia variegata). Emerg Infect Dis. 2002;8:1497–500.

    Article  Google Scholar 

  16. 16.

    Dantas-Torres F, Chomel BB, Otranto D. Ticks and tick-borne diseases: a One Health perspective. Trends Parasitol. 2012;28:437–46.

    Article  Google Scholar 

  17. 17.

    Stuen S. Anaplasma phagocytophilum – the most widespread tick-borne infection in animals in Europe. Vet Res Commun. 2007;31:78–9.

    Article  Google Scholar 

  18. 18.

    Yabsley MJ, Norton TM, Powell MR, Davidson WR. Molecular and serologic evidence of tick-borne Ehrlichiae in three species of lemurs from St. Catherines Island, Georgia, USA. J Zoo Wildl Med. 2004;35:503–9.

    Article  Google Scholar 

  19. 19.

    Miller DS, Sauther ML, Hunter-Ishikawa M, Fish K, Culbertson H, Cuozzo PF, et al. Biomedical evaluation of free-ranging ring-tailed lemurs (Lemur catta) in three habitats at the Beza Mahafaly Special Reserve, Madagascar. J Zoo Wildl Med. 2007;38:201–16.

    Article  Google Scholar 

  20. 20.

    Dzięgiel B, Adaszek Ł, Carbonero A, Łyp P, Winiarczyk M, Dębiak P, et al. Detection of canine vector-borne diseases in eastern Poland by ELISA and PCR. Parasitol Res. 2016;115:1039–44.

    Article  Google Scholar 

  21. 21.

    Adaszek Ł, Górna M, Skrzypczak M, Buczek K, Balicki I, Winiarczyk S. Three clinical cases of Anaplasma phagocytophilum infection in cats in Poland. J Feline Med Surg. 2013;15:333–7.

    Article  Google Scholar 

Download references


Not applicable.


The authors received no specific funding for this work.

Author information




AW, JZ and OT performed the clinical evaluation, collected the sample and administered the treatment. DW was involved in the hematological and biochemical analysis. LA performed PCR analysis. LA, MK, MS and SW were involved in the data interpretation. LA drafted the manuscript and MK and SW critically read and edited the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Marcin Kalinowski.

Ethics declarations

Ethics approval and consent to participate

This study did not require the approval of an ethical committee since it is a case report and samples used were surplus material from the diagnostic tests.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Adaszek, Ł., Wilczyńska, A., Ziętek, J. et al. Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland. BMC Vet Res 17, 118 (2021).

Download citation


  • Anaplasma phagocytophilum
  • Ring-tailed lemur
  • Vector-borne disease
  • Poland