In vitro isolation and identification of the first Neospora caninum isolate from European bison (Bison bonasus bonasus L.)
Introduction
Neospora caninum is an important an obligate intracellular protozoan that causes abortion and economic losses in the cattle industry. The parasite has also been detected in other livestock species, companion animals and wildlife animals (Dubey, 2003, Gondim, 2006). There is much current interest in wild animal reservoirs for the parasite.
Neospora caninum infection was confirmed in many wild ruminant species such as black-tailed deer (Odocoileus hemionus columbianus), white-tailed deer (O. virginianus), moose (Alces alces), mule deer (O. hemionus hemionus), caribou (Rangifer tarandus), American bison (Bison bison), chamois (Rupicapra rupicapra), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) (Dubey et al., 1999, Ferroglio et al., 2003, Dubey, 2003, Dubey and Thulliez, 2005, Wolf et al., 2005, Gondim, 2006, Almeria et al., 2007). In Europe, the presence of the parasite has been serologically shown in free-living and captive red deer (C. elaphus), roe deer (C. capreolus), fallow deer (Dama dama), sika deer (C. nippon) and also in free-living and captive European bison (Bison bonasus bonasus L.) (Ferroglio and Rossi, 2001, Cabaj et al., 2005, Almeria et al., 2007, Bartova et al., 2007, Goździk et al., 2010). European bison (B. bonasus bonasus L.) is the largest herbivorous animal in Europe. Historically, the species was distributed throughout western, central, and south-eastern Europe and the Caucasus. In 1951, the Lowland (Białowieża) bison were reintroduced successfully on wild grounds of the Polish part of the Białowieża Forest. In 1966 the Union of International for the Conservation of the Nature and Natural Resources (IUCN) formed the Red List of Threatened Species which classified the bison as the endangered species. European bison in Białowieża Forest play an important role in the restitution program and protection of the species.
The aim of the study was to monitor European bison in terms of the presence of antibodies against N. caninum and to isolate the parasite and to characterize it using PCR techniques and sequencing analysis.
Section snippets
Animals and source of samples
The Białowieża Forest (the last ancient primeval woodland in Europe) is a contiguous forest complex covering 1500 km2. The western part this complex is located in Poland and covers 650 km2. The eastern (Belarusian) part is larger covering 875 km2. The Forest extends for 55 km in the east-west direction and 51 km from north to south. The area is delineated by the coordinates 23°31′–24°21′E and 52°29′–52°37′N (Sokołowski, 2004).
In this study, 23 blood samples were collected from the jugular vein
ELISA and Western blot
In the population of 23 European bison eliminated in Białowieża Forest in 2009, three of them showed a strong positive response to N. caninum (13%). These positive sera were tested also by Western blot. The results confirmed the presence of antibodies to Neospora tachyzoites antigens. The antibody response was directed against a wide range proteins (9.5, 17, 21, 27, 31, 36.5, 38,40,43,47, 48.5, 53.5 and 58 kDa). The most heavily stained bands had molecular weights of 9.5, 17, 27 and 58 kDa.
The
Discussion
Previously, Cabaj et al. (2005) reported for the first time the presence of specific antibodies in 13 (7.3%) among 320 examined European bison (B. bonasus bonasus L.) living in free and fenced areas in Białowieża Forest. The first record of antibodies against N. caninum in European bison was found in a 16 years old cow, lived in the Białowieża Forest in freedom, and shot on December 14th, 1988 (Cabaj et al., 2005). It is worth noticing, that the earliest detection of European bison positive
Acknowledgements
Grateful thanks are due to Dr. Katarzyna Goździk for help in sequence analysis of the Polish bison isolate N. caninum.
The research is financed by The Ministry of Sciences and Higher Education, Poland, grant no. N303 062 32/2263.
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2018, Parasitology InternationalIsolation of Neospora caninum from kidney and brain of a bovine foetus and molecular characterization in Brazil
2018, Experimental ParasitologyCitation Excerpt :Parasites were also isolated from brain samples of the same foetus, and the tachyzoites were first observed on day 20 post-inoculation with brain homogenate. The time between the inoculation of Vero cells and the first visible tachyzoites in the culture probably depends on the number of tachyzoites in the sample (Bien et al., 2010). In the present study, the abortion took place in the second trimester of the gestation, suggesting that the parasite load was high (Collantes-Fernandez et al., 2005).
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2015, International Journal for Parasitology: Parasites and WildlifeCitation Excerpt :The protozoan cyst was considered an incidental finding. Neospora caninum antibodies and/or DNA have been detected in the following free-ranging nondomestic artiodactyls: white-tailed deer (Odocoileus virginianus) (Ab) (Dubey et al., 1999, 2009; Lindsay et al., 2002; Gondim et al., 2004a; Anderson et al., 2007; Gutierrez-Exposito et al., 2012; Olamendi-Portugal et al., 2012), black-tailed deer (Odocoileus hemionus columbianus) (Ab, DNA) (Woods et al., 1994; Dubey et al., 2008), mule deer (Odocoileus hemionus hemionus) (Ab) (Dubey et al., 2008; Myers et al., 2014), pampas deer (Ozotoceros bezoarticus) (Ab) (Tiemann et al., 2005a), roe deer (Capreolus capreolus) (Ab) (Bartova et al., 2006; Almeria et al., 2007; Panadero et al., 2010; De Craeye et al., 2011; Malmsten et al., 2011; Candela et al., 2014), fallow deer (Dama dama) (Ab) (Bartova et al., 2006; Marco et al., 2008), red deer (Cervus elaphus and C. elephus yarkandensis) (Ab) (Ferroglio and Rossi, 2001; Bregoli et al., 2006; Almeria et al., 2007; Gozdzik et al., 2010; Billinis, 2013), elk (Cervus canadensis) (Ab) (Gutierrez-Exposito et al., 2012), moose (Alces alces) (Ab) (Dubey and Thulliez, 2005; Stieve et al., 2010; Malmsten et al., 2011; Gutierrez-Exposito et al., 2012; Moskwa et al., 2014), caribou (Rangifer tarandus) (Ab) (Dubey and Thulliez, 2005; Stieve et al., 2010; Gutierrez-Exposito et al., 2012), North American bison (Bison bison) (Ab) (Dubey and Thulliez, 2005), European bison (Bison bonasus bonasus) (Ab) (Cabaj et al., 2005; Bien et al., 2010), African buffalo (Syncerus caffer) (Ab) (Ferroglio et al., 2003), Thompson's gazelle (Eudorcas thomsonii) (Ferroglio et al., 2003), impala (Aepyceros melampus) (Ab) (Ferroglio et al., 2003), eland (Taurotragus oryx) (Ab) (Ferroglio et al., 2003), chamois (Rupicapra rupicapra) (Ab, DNA) (Ferroglio and Rossi, 2001; Bregoli et al., 2006; Gaffuri et al., 2006; De Craeye et al., 2011), Alpine ibex (Capra ibex) (Ab) (Bregoli et al., 2006), Spanish ibex (Capra pyrenaica hispanica) (Ab) (Garcia-Bocanegra et al., 2012), European mouflon (Ovis musimon) (Bartova et al., 2006), Barbary sheep (Ammotragus lervia) (Ab) (Almeria et al., 2007), musk ox (Ovibos moschatus) (Ab) (Dubey and Thulliez, 2005), wild boar (Sus serofa) (Ab) (Bartova et al., 2006; Almeria et al., 2007), and warthog (Phacochoerus aethiopicus) (Ab) (Ferroglio et al., 2003). Neospora caninum antibodies have also been detected in the following captive nondomestic artiodactyl species: Axis deer (Axis axis) (Basso et al., 2014), Eld's deer (Cervus eldi siamesis) (Dubey et al., 1996), Thorold's deer (Cervus albirostris) (Sedlak and Bartova, 2006), eastern elk (Cervus elaphus canadensis) (Sedlak and Bartova, 2006), Vietnam sitka deer (Cervus Nippon pseudaxis) (Bartova et al., 2006; Sedlak and Bartova, 2006), Brocket deer (Mazama spp.) (Tiemann et al., 2005b), Pere David's deer (Elaphurus davidianus) (Sedlak and Bartova, 2006), antelope (Tragelaphus iberbis) (Peters et al., 2001), blackbuck (Antilope cervicapra) (Sedlak and Bartova, 2006), lechwa (Kobus leche) (Sedlak and Bartova, 2006), African buffalo (Syncerus caffer caffer) (Sedlak and Bartova, 2006), eland (Taurotragus oryx) (Sedlak and Bartova, 2006), European bison (Bison bonasus bonasus) (Sedlak and Bartova, 2006), sitatunga (Tragelaphus spekei gratus) (Sedlak and Bartova, 2006), and vicuña (Vicugna vicugna mensalis) (Risco-Castillo et al., 2014).