RESEARCH PAPER
Comparison of the detection efficiency of haemoparasite DNA in blood and faecal samples – the way to eco-epidemiological studies
More details
Hide details
1
University of Warsaw, Poland
Corresponding author
Anna Bajer
University of Warsaw, Miecznikowa 1, 02-096, Warszawa, Poland
Ann Agric Environ Med. 2019;26(4):538-543
KEYWORDS
TOPICS
ABSTRACT
Introduction and objective:
It is easier and non-invasive to obtain faecal samples compared with blood samples. Molecular techniques may enable detection of parasites even in tiny amounts of blood-containing faeces. We aimed to compare the sensitivity of detection of three Babesia species and Hepatozoon canis in blood and faecal samples, including samples
derived from naturally infected hosts.
Material and methods:
Three groups were involved: 1) Nine BALB/c mice infected with Babesia microti sampled during
acute (n=3), post-acute (n=3) and chronic phases of infection (n=3); 2) Eight dogs with symptoms of babesiosis; 3) Six red foxes infected with B. vulpes, one fox infected with B. canis, four foxes infected with H. canis. Genomic DNA was extracted from blood and faeces by use of commercial kits and amplified with genus-specific primers in one-step or nested PCR reactions. Selected PCR products were sequenced.
Results:
No positive results for faecal samples were obtained from H. canis-positive foxes in contrast to Babesia spp.
infections. Positive results from PCRs were obtained for all BALB/c mice (100%), five dogs (62.5%) and four of seven foxes (57.1%). Successful sequencing was obtained for six selected murine samples (B. microti), four canine samples (B. canis) and for one fox sample (B. vulpes). The success of B. microti detection in murine faecal samples from acute, post-acute and chronic phases was identical (100%).
Conclusions:
Detectability of Babesia spp. infections was lower in naturally infected dogs and foxes, compared to
experimentally infected mice. Detection of DNA in faecal samples can be useful in the detection of Babesia infection in
populations from which blood samples are hard to obtain, but due regard must be given to the possibility that prevalence of infection may be severely underestimated.
ACKNOWLEDGEMENTS
The authors express their thanks to the owners of the dogs
for their kind cooperation. Professor Jerzy M. Behnke at the
University of Nottingham, UK, undertook a critical review
of the manuscript. The study was partially supported by
the National Science Centre (NCN), grant Sonata Bis No.
2014/14/E/NZ7/00153, who had no role in the design of the
study and collection of samples, analysis and interpretation
of data and, or in the writing of the manuscript.
REFERENCES (32)
1.
Kloch A, Bednarska M, Bajer A. Intestinal macro- and microparasites of wolves (Canis lupus L.) from north-eastern Poland recovered by coprological study. Ann Agric Environ Med. 2005; 12:237–245.
2.
Paziewska A, Bednarska M, Nieweglowski H, Karbowiak G, Bajer A. Distribution of Cryptosporidium and Giardia spp. in selected species of protected and game mammals from North-Eastern Poland. Ann Agric Environ Med. 2007; 14:265–270.
3.
Jirku M, Votypka J, Petrzelkova KJ, Jirku-Pomajbikova K, Kriegova E, Vodicka R, et al. Wild chimpanzees are infected by Trypanosoma brucei. Int J Parasitol Parasites Wildl. 2015; 4:277–282.
4.
Keita AK, Fenollar F, Socolovschi C, Ratmanov P, Bassene H, Sokhna C, et. al. The detection of vector-borne-disease-related DNA in human stool paves the way to large epidemiological studies. Eur J Epidemiol. 2015; 30:1021–1026.
5.
Siregar JE, Faust CL, Murdiyarso LS, Rosmanah L, Saepuloh U, Dobson AP, Iskandriati D. Non-invasive surveillance for Plasmodium in reservoir macaque species. Malar J. 2015; 14:404.
6.
Zygner W. Arthropod-borne parasitoses which may threaten health of dogs travelling into Mediterranean area and Portugal. Part II. Babesiosis, theileriosis and hepatozoonosis. Życie Weterynaryjne 2006; 81:595–603.
7.
Baric Rafaj R, Kules J, Selanec J, Vrkic N, Zovko V, Zupancic M, et al. Markers of coagulation activation, endothelial stimulation, and inflammation in dogs with babesiosis. J Vet Intern Med. 2013; 27:1172–1178.
8.
Weerakoon KG, McManus DP. Cell-free DNA as a diagnostic tool for human parasitic infections. Trends Parasitol. 2016; 32:378–391.
9.
Kaiser M, Lowa A, Ulrich M, Ellerbrok H, Goffe AS, Blasse A, et al. Wild chimpanzees infected with 5 Plasmodium species. Emerg Infect Dis. 2010; 16:1956–1959.
10.
Liu W, Li Y, Learn GH, Rudicell RS, Robertson JD, Keele BF, et al. Origin of the human malaria parasite Plasmodium falciparum in gorillas. Nature 2010; 467:420–425.
11.
Jirku M, Pomajbikova K, Petrzelkova KJ, Huzova Z, Modry D, Lukes J. Detection of Plasmodium spp. in human faeces. Emerg Infect Dis. 2012; 18:634–636.
12.
Liu W, Li Y, Shaw KS, Learn GH, Plenderleith LJ, Malenke JA, et al. African origin of the malaria parasite Plasmodium vivax. Nat Commun. 2014; 5:3346.
13.
Keele BF, Van Heuverswyn F, Li Y, Bailes E, Takehisa J, Santiago ML, et al. Chimpanzee reservoirs of pandemic and nonpandemic HIV-1. Science 2006; 313:523–526.
14.
El Khechine A, Henry M, Raoult D, Drancourt M. Detection of Mycobacterium tuberculosis complex organisms in the stools of patients with pulmonary tuberculosis. Microbiology 2009; 155:2384–2389.
15.
Hornok S, Estok P, Kovats D, Flaisz B, Takacs N, Szoke K, et al. Screening of bat faeces for arthropod-borne apicomplexan protozoa: Babesia canis and Besnoitia besnoiti-like sequences from Chiroptera. Parasit Vectors 2015; 8:441.
16.
Welc-Faleciak R, Bajer A, Bednarska M, Paziewska A, Sinski E, Long term monitoring of Babesia microti infection in BALB/c mice using nested PCR. Ann Agric Environ Med. 2007; 14:287–290.
17.
Bednarska M, Bajer A, Drozdowska A, Mierzejewska EJ, Tolkacz K, Welc-Faleciak R. Vertical Transmission of Babesia microti in BALB/c Mice: Preliminary Report. PLoS One 2015; 10(9):e0137731.
18.
Bonnet S, Jouglin M, L’Hostis M, Chauvin A. Babesia sp. EU1 from roe deer and transmission within Ixodes ricinus. Emerg Infect Dis. 2007; 13:1208–1210.
19.
Bonnet S, Jouglin M, Malandrin L, Becker C, Agoulon A, L’Hostis M, Chauvin A. Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique. Parasitology 2007; 134:197–207.
20.
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.
21.
Mitkova B, Hrazdilova K, D’Amico G, Duscher GG, Suchentrunk F, Forejtek P, et al. Eurasian golden jackal as host of canine vector-borne protists. Parasit Vectors 2017; 10:183.
22.
Qurollo BA, Archer NR, Schreeg ME, Marr HS, Birkenheuer AJ, Haney KN, et al. Improved molecular detection of Babesia infections in animals using a novel quantitative real-time PCR diagnostic assay targeting mitochondrial DNA. Parasit Vectors 2017; 10:128.
23.
Baneth G, Florin-Christensen M, Cardoso L, Schnittger L. Reclassification of Theileria annae as Babesia vulpes sp. nov. Parasit Vectors 2015; 8:207.
24.
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.
25.
Hodzic A, Alic A, Fuehrer HP, Harl J, Wille-Piazzai W, Duscher GG. A molecular survey of vector-borne pathogens in red foxes (Vulpes vulpes) from Bosnia and Herzegovina. Parasit Vectors 2015; 8:88.
26.
Hodzic A, Mrowietz N, Cezanne R, Bruckschwaiger P, Punz S, Habler VE, et al. Occurrence and diversity of arthropod-transmitted pathogens in red foxes (Vulpes vulpes) in western Austria, and possible vertical (transplacental) transmission of Hepatozoon canis. Parasitology 2018; 145:335–344.
27.
Miterpakova M, Komjati-Nagyova M, Hurnikova Z, Vichova B. Retrospective molecular study on canine hepatozoonosis in Slovakia – Does infection risk for dogs really exist? Ticks Tick Borne Dis. 2017; 8:567–573.
28.
Majlathova V, Hurnikova Z, Majlath I, Petko B. Hepatozoon canis infection in Slovakia: imported or autochthonous? Vector Borne Zoonotic Dis. 2007; 7:199–202.
29.
Matijatko V, Torti M, Schetters TP. Canine babesiosis in Europe: how many diseases? Trends Parasitol. 2012; 28:99–105.
30.
Kubelova M, Tkadlec E, Bednar M, Roubalova E, Siroky P. West-to-east differences of Babesia canis canis prevalence in Dermacentor reticulatus ticks in Slovakia. Vet Parasitol. 2011; 180:191–196.
31.
Foldvari G, Hell E, Farkas R. Babesia canis canis in dogs from Hungary: detection by PCR and sequencing. Vet Parasitol. 2005; 127:221–226.
32.
Koneval M, Miterpakova M, Hurnikova Z, Blanarova L, Vichova B. Neglected intravascular pathogens, Babesia vulpes and haemotropic Mycoplasma spp. in European red fox (Vulpes vulpes) population. Vet Parasitol. 2017; 243:176–182.