A coprological survey of parasitic fauna firstly in Wild Far Eastern Leopard (Panthera pardus orientalis)

BackgroundThe Amur leopard, one of nine recently recognized subspecies of leopard, is still most threatened in a stochastic procession of extinction. The potential harmless to the conservation of the Amur leopard originating from the disease is in need of urgent attention. Unfortunately, the research on the potential risk to Amur leopard caused by disease is rare. When the parasites were concerned, even the elementary data, such as parasitic fauna, are absent. Our aim in the study is to accumulate the knowledge of it for a better comprehension. Results: There are 7 parasite species, including 3 nematodes (Toxocara cati, capillarid-type parasite, and Metastrongyloidea-type parasite), 2 cestodes (Spirometra sp. and Taenia sp.), 1 trematode (Paragonimus sp.), and 1 protozoa (Cystoisospora felis), were found in this research. The Toxocara cati was highest frequent occurrence, followed by Spirometra sp. Conclusion: The Amur leopard was infected by seven parasites firstly reported.


Discussion
The Toxocara cati is a worldwide parasite of felids, and it is very popular in the both captured and wild Siberian tigers [9][10][11]. Our research indicates that the Toxocara cati also is a common parasite in wild Amur leopard with a high frequency of 61.9%. In general, the Amur tiger, nearly completely sympatric with Amur leopard, could infect two kinds of roundworm, Toxocara cati and Toxascaris leonine. For the captured tiger, both of them are very popular, whereas Toxascaris leonine is rare in the wild Amur tigers [9][10][11]. The situation of Amur leopard is little difference in Amur leopard as there no Toxascaris leonine was found in the wild population in current study.  [17]. Eucoleus aerophilus is a globally distributed parasite among various wild carnivorous mammals, and resides embedded in host's epithelium of the trachea, bronchi and bronchioles of the lungs. Usually, the pathogenicity of it is considered subclinical with respiratory distress coughing and wheezing, may be the clinical presentations in heavily infected hosts [15]. Aonchotheca putorii is a parasitic nematode of the stomach and small intestines of many wild mammals, and causes severe gastric, associated with a gastric ulcer and secondary anemia [16]. What special species the parasite is need a molecular identification in future.  [34] .
Amur leopard, a big size cat, Holarctic felid, preys on a wide range of mammals, but it's overwhelming majority of diets were roe deer (up to 66%), wild boars (up to 8%), Siberian musk deer (up to 9%), and sika deer (up to 6%)[7]. Therefore, Amur leopard was involved in the life cycles of Taenia sp is not out anticipation. Although it could be concluded that Taenia worms infected the Amur leopard based on presenting the distinctly Taenia eggs, developed larva with 3 pairs of hooks, and surrounded by a thick and striated shell, however, it is difficult to discriminate the Taenia species only based on the morphological characteristics of the eggs as all species of Taenia genus share those special characteristics.
The Paragonimus spp., lung flukes, are trematodes that parasitize the lungs of mammal animals like 6 Carnivores, the major definitive host.  [37]. The latter route seems to be more important in Amur leopard, as it always take the wild boar, sika deer and roe deer as food other than crustaceans There are about fifty nominal Paragonimus species, meanwhile, over half of them have been found in China, one of the major endemic foci of paragonimiasis in the world. Two of them, P. westermani and P. skrjabini, known zoonotic parasites, are more commonly or focused in China. Although most of Paragonimus spp. occur in tropical and subtropical regions, however, two species, P. westermani and P. ohirai, extend far into temperate latitudes of China [35]. The P. westermani , the only one, converges the habitats of the Amur leopards. Although twenty of them were found within the cat, while only seven of them, including P. westermani, were found in wild felines [35]. For P. westermani, it was initially recognized from a Bengal Tiger at the Amsterdam zoological gardens, which migrated from Asia. Thereafter, other seven feline hosts, including felis catus, Catopuma temminckii, Neofelis nebulosa, Panthera pardus, Prionailurus bengalensis, Prionailurus planiceps, and Prionailurus rubiginosus, found could be infected by P. westermani [35]. The trematode of Amur leopard was easily identified as paragonimus with the features of the eggs, the distantly shoulders or opercular ridges which are useful for differentiating Paragonimus eggs from the large operculate eggs of other trematodes [38]. For the specific Paragonimus species discrimination, with a high degree certainty, it was the P. westermani based on the epidemiological data, such as the parasite's endemic region and host species recorded in the past. However, it is not sufficient to deny the possibility of the other Paragonimus spp. without molecular evidence.
There are only two coccidian species, Cystoisospora felis (syn. Isopora felis) and C. rivolta (syn. Isopora rivolta), in cats. The typical symptom of Cystoisospora was diarrhea in kittens, occasionall mortality for C. Felis [39]. The egg size of C. felis (32-53×26-43 μm) is much bigger than C. rivolta 7 (18-25 x 16-23 μm) [39], so it was certainly identified that the coccidian species from Amur leopard as the C. felis based on the eggs sizes. Until now, with the Amur leopard together, those wild felines, Amur leopard as their population density are much lower that domestic felines. Therefore, paratenic host must play an important role in transmission of C. felis. Actually, based on the results of bioassay studies, the animals like dog, cattle, pig, mice, and rabbit may act as paratenic host for Cystoisospora has been discussed in the past [39].

Conclusion
The Amur leopard were infected by seven parasites firstly reported.

Methods 8
The research was carried in the Northeast of China, and most samples were collected in the Wangqing National Nature Reserve (Fig.2). After permission was granted by local government, 42

Ethics approval and consent to participate
Collecting the feces samples from the wild Amur leopard was approved by the local government agents.

Availability of data and materials
All the data and materials used in this report are included in the manuscript.

Competing interests
The authors declare that they have no competing interests.

Funding
This study was supported by the following Grants: National Key Research and Development Program    The picture showed the site of the samples were collected, it was made by one author Dr Zhiwei Peng.