Conceived and designed the experiments: Balantidiasis is considered a neglected zoonotic disease with pigs serving as reservoir hosts. However, Balantidium coli has been recorded in many other mammalian species, including primates. Here, we evaluated the genetic diversity of B. We analyzed 49 isolates of ciliates from fecal samples originating from 11 species of captive and wild primates, domestic pigs and wild boar. The phylogenetic trees were computed using Bayesian inference and Maximum likelihood.
Balantidium entozoon from edible frog and Buxtonella sulcata from cattle were included "Nigeria och kamerun vm klara 3" the analyses
Nigeria och kamerun vm klara 3 the closest relatives of B. Thus, we propose a new genus Neobalantidium for the homeothermic clade. Moreover, several isolates from both captive and wild primates excluding great apes clustered with B. The cysts of Buxtonella and Neobalantidium are morphologically indistinguishable and the presence of Buxtonella -like ciliates in primates opens the question about possible occurrence of these pathogens in humans.
Balantidium coli is a pathogenic ciliate occurring in various hosts, including primates. Balantidiasis is considered a neglected disease with zoonotic potential and it is associated with pigs as reservoirs. Although it is considered to be rare, a high prevalence of B. The infection can cause symptoms ranging from mild diarrhea to fulminating dysentery. Recently, balantidiasis has appeared to be a serious problem in immunocompromised persons.
Although a chapter about balantidiasis is included in almost every parasitology textbook, many aspects of the occurrence of these ciliates in mammals, including humans, remain unknown. As balantidiasis can pose a serious health problem, a better understanding of this organism has important practical implications. We performed an extensive evaluation of genetic diversity in B. We find that the diversity of the cyst-forming ciliates in non-human primates is broader than expected.
This discovery poses novel questions about the real spectrum of intestinal ciliates occurring in humans, and might explain unknown factors in the epidemiology and pathogenicity of ciliate infections.
Balantidiidae is a cosmopolitan ciliate colonizing the intestine of many mammalian hosts. However, domestic pigs and wild boars are considered to be the principal host and major reservoir .
Balantidiasis is considered a zoonotic disease and human clinical cases in developed countries were typically associated with close contact with pigs in the past .
Nowadays, localities with a high prevalence of B. Apart from humans, B. The clinical importance of B. Presently, human populations living in close proximity with domestic pigs are naturally resistant and mostly without any clinical manifestation . However, the infection can cause disease, with symptoms ranging from mild diarrhea to fulminating dysentery.
On rare occasions these organisms may also invade other which is more frequently observed in immunocompromised individuals afflicted with AIDS or leukemia .
The Balantidium taxonomy is somewhat controversial due to the pleomorphism of its trophozoites  and range of its hosts. Balantidium coli observed in dysenteric patients was originally described as Paramecium coli by Malmstein .
A great majority of the taxa included in this genus were isolated from amphibian, fish or insect hosts . In mammals, all balantidia are currently referred to as B. The broad synonymy of B. Apparently, trophozoite morphology alone is insufficient for taxonomical purposes. The worldwide distribution of B. Currently, fairly extensive and congruent molecular phylogenies have been obtained based on small ribosomal subunits of free-living and commensal ciliates in recent years .
The SSrRNA is a gene of taxonomic relevance at the genus level and, nowadays, is broadly used for taxonomic studies in combination with morphological features [e. Currently, only few SSrRNA sequences of Balantidium are available in the GenBank and their comparison across various hosts, pigs, ostriches and gorillas, has revealed little variability  ; however, the sequence of B.
Recently, the molecular diversity of B. Such ambiguous polymorphism questions the applicability of internal transcribed spacers as markers for analyzing the intraspecific genetic variability of B. Our previous research on the occurrence of B.
Although balantidiasis is a neglected disease, molecular diversity of Balantidium in humans has not yet been addressed. Based on the samples from captive and wild non-human primates, together with comparative material from domestic pigs in Europe and tropical Nigeria och kamerun vm klara 3, we performed an extensive evaluation of the genetic diversity of B. Our study was completely non-invasive including only examination of fecal samples. The only invasive sampling was the use of cloacal lavage in edible frogs.
This material was collected prior to our research during the field course for undergraduate students. As such, it was approved by Administratia Rezervatiei Biosferei Delta Dunarii for details about the permits and collaborating authorities see Text S1. Fecal samples from captive primates were obtained in collaboration with i several European Zoos and ii African sanctuaries during routine diagnostic check-ups. Every facility was provided with the results, which they later used for antiparasitic control.
Nigeria och kamerun vm klara 3 European countries, our research was approved by particular Zoos which follow their respective national animal care regulations or guidelines. To our knowledge, and based on long-term research on primate parasites, such non-invasive fecal sampling has no other regulations in the EU. Thus, this study complies with individual national animal care regulations or guidelines. Other material included fecal samples from domestic pigs containing B.
In the wild, fecal samples were obtained during distance tracking of the animals. Fecal samples of edible frogs were obtained by cloacal lavage. All samples were analyzed coproscopically using Sheather's flotation and Merthiolate-iodine-formaldehyde MIFC sedimentation .
The ITS1 - 5. The amplicons were purified using the Qiagen extraction gel kit. Sequencing of these amplicons was performed by Macrogen, Korea.
Each sample was sequenced in both directions and the sequence contigs were assembled using the program ChromasPro, version 1. BLAST analyses were performed for all new sequences.
Both alignments were manually edited in BioEdit  and ambiguously aligned. In the case of ITS sequences, the ambiguously aligned regions were omitted from further analyses.
Alignments were analyzed individually and also in a combined data set. Branching support was estimated from non-parametric bootstrap replicates using the above mentioned software Nigeria och kamerun vm klara 3 conditions.
Bayesian topologies were estimated using Phylobayes 3. The latter is a combination of the empirical mixture model of 40 profiles C40 sharing exchange rates as defined by the GTR model. For all data sets, two independent chains were run until they converged i. Bayesian posterior probabilities PP provide branching support for the resulting topology only branches with a PP higher than 0.
The possible hypotheses for the evolutionary history of the genus Balantidium were tested using the approximately unbiased AU test  on the SSrRNA gene dataset. Alternative topologies of the in-group Fig. The site-specific log-likelihood scores of the resulting trees were obtained by the same software and further analyzed using the AU-test as implemented in CONSEL .
The numbers above the branches represent Maximum likelihood bootstrap supports as computed from replicates. The scale bar represents 10 changes per positions. The tree is complemented by an AU topology test, with all tested topologies shown below the main tree topologies shown in reduced form. New sequences are marked with a star. The sequences of both above-mentioned markers obtained in this study from each host are available in GenBank under the accession numbers JQ, JQ Table 1 — 3.
This tree was rooted using Epispathidium papilliferum DQ as an outgroup. The maximum likelihood RAxML with GTR model topology of our resulting tree shows that the subclass Trichostomatia is a monophyletic and highly supported group divided into two major clades. However, the order Vestibuliferida appears to be paraphyletic in its origin Fig. Clade I includes all the sequences of Balantidium and is further divided into two sub-clades: Although the topology inferred from SSrRNA genes is unstable, it is very likely that the genus Balantidium is polyphyletic.
Such topology is strongly preferred by the AU test Fig. This "Nigeria och kamerun vm klara 3" displays internal polytomy, obviously due to the presence of a high number of similar sequences. Several ciliates from primates agile mangabeys: JQ; the hamadryas baboon: JQ formed a separate clade that branched together with sequences of B. However, such a relationship is only very weakly supported by ML trees; trees computed by PhyloBayes placed B.
The phylogenetic trees based on the ITS This sequence data set includes our 52 sequences and reference sequences from GenBank that are comprised of five isolates of T.
The resulting ML tree topology shows three major groups: The majority of the supposed B. The A-group includes 21 of our ciliate sequences "Nigeria och kamerun vm klara 3" 11 reference sequences from captive great apes, two sequences from humans, one from wild boar, two from ostriches and six sequences from domestic pigs.
The B-group contains 10 newly obtained sequences from ciliates from domestic pigs from different topodemes and four reference sequences 3 isolates from ostriches: All obtained sequences originating from domestic pigs belong to type of genetic variant B. Importantly, the sequences of all 17 isolates of ciliates obtained from primates other than great apes and humans, together with B. This group of isolates from primates is further divided into two strongly supported sub-groups that may represent a type of genetic variants.
In this analysis, B. The standard approach to measuring medicine prices and . Douglas Ball, Andrew Chetley and Klara Tisocki who drafted specific chapters in the .
mark- ups; the Government of Nigeria, which is drafting a medicines policy, based on. AIMs-based estimates for Senegalese and Nigerian samples (far right).
3) Niger, Nigeria, and Cameroon, 4) Guinea-Bissau, Mali, Senegal. Mar; 7(3): e 3. Areán VM, Koppisch E () Balantidiasis. parasites in crop raiding and wild foraging Papio anubis in Nigeria.