Neobalantidium coli: First molecular identification from the Eurasian wild boar, Sus Scrofa in Bushehr Province, Southwestern Iran.

Balantidium coli is a common parasite of pig and wild boars (Sus scrofa) which can infect humans and several species of mammals. This study aimed to determine the genotype of Balantidium isolated from Eurasian wild boars in Bushehr province, Southwestern Iran. Twenty-five faecal samples, originating from 25 wild boars captivated in our previous study, were processed. DNA was extracted from the faecal samples and PCR-amplified, targeting an ITS1-5.8s-rRNA-ITS2 region of Balantidium genome. PCR product was purified from the gel, and sequenced. BLAST analysis was performed in order to compare our isolates with other previously reported ones. A phylogenetic tree was constructed, using MegaX software, to find out the phylogenetic diversity of the isolates. With PCR it was possible to detect Balantidium DNA in the faecal samples of 13 out of 25 (52%) of the wild boars. BLAST analysis of seven isolates revealed that the isolates belong to the newly introduced genus Neobalantidium coli. Sequences of three isolates were deposited in the GenBank. Moreover, molecular analysis revealed six areas of nucleotide differences within the isolates and nine areas of difference between the sequences obtained in this study and those available in the GenBank. Phylogenetic analysis revealed that the sequences of isolates of this study have up to 2.2% dissimilarity from those published in the GenBank. The findings of this study, for the first time, revealed that some of the isolates of Balantidium originating from wild boars in Southwestern Iran belonged to the N. coli.

First molecular identification of Buxtonella ciliates from captive-bred mangabeys (Cercocebus torquatus) from China.

Buxtonella species are large cyst-forming ciliates that infect ruminants and monkeys, and are morphologically similar to Balantidium coli ciliates that infect pigs, humans, monkeys, and other animals. In this study, we isolated spherical cysts of ciliates that were similar to those of Balantidium and Buxtonella species within collared mangabeys (Cercocebus torquatus) from the Wangcheng Zoo of Luoyang in the Henan Province of central China. The cysts were further identified and designated as belonging to the Buxtonella monkey genotype based on molecular analyses of 18S rRNA, 5.8S rRNA, and ITS1-5.8S rRNA-ITS2 genetic markers. To our knowledge, this is the first report of Buxtonella monkey genotype within monkeys in China. These results will help clarify the classification of species of cyst-forming ciliate infections in monkeys.

Balantidium grimi n. sp. (Ciliophora, Litostomatea), a new species inhabiting the rectum of the frog Quasipaa spinosa from Lishui, China.

Balantidium grimi n. sp. is described from the rectum of the frog Quasipaa spinosa (Amphibia, Dicroglossidae) from Lishui, Zhejiang Province, China. The new species is described by both light microscopy (LM) and scanning electron microscopy (SEM), and a molecular phylogenetic analysis is also presented. This species has unique morphological features in that the body shape is somewhat flattened and the vestibulum is "V"-shaped, occupying nearly 3/8 to 4/7 of the body length. Only one contractile vacuole, situated at the posterior body, was observed. The phylogenetic analysis based on SSU-rDNA indicates that B. grimi groups together with B. duodeni and B. entozoon. In addition, the genus Balantidium is clearly polyphyletic.

Diversity and prevalence of gastrointestinal parasites in two wild Galago species in Gabon.

In this study, we characterize the diversity and estimated infection levels of gastrointestinal parasites circulating in two galago species, Galago demidoff and G. thomasi in two sites situated in the Southeastern forests of Gabon. Our study reveals that eleven parasites including nine helminthes (Ascaris spp., Ankylostoma spp., Dicrocoelium spp., Gongylonema spp., Oesophagostomum spp., Lemuricola spp., Strongyloides spp. Trichostrongylus spp. and Trichuris spp.) and two protozoans (Balantidium spp. and Entamoeba spp.) may infect Galago spp. with high infection rates. The results show that: a very similar parasite spectrum is found in both host species; all the taxa identified were previously observed in other Primate species and/or Man. They also show that age, gender and forest type may influence infection rates and/or parasite diversity found in a particular host and/or geographic area.

The First Case of Diarrhoea in Tibetan Sheep, Ovis aries, Caused by Balantidium coli in the Qinghai Tibetan Plateau Area, China.

This study was carried out to determine the pathogen-causing diarrhoea in sheep Ovis aries in the Qinghai Tibetan Plateau Area, China. A trophozoite was identified as species of ciliate alveolates infecting the sheep based on morphological characteristics examined by microscope. It was mostly spherical, colourless and transparent, with many vesicles. Macronucleus and contractile vacuoles could not be distinguished. Size of the trophozoite was 80-180 × 70-150 µm and its surface was covered with cilia. Molecular analysis based on sequences of 18S rRNA and ITS genes confirmed the ciliate species as Balantidium coli. According to the literature, there have been many epidemiological investigations of B. coli infection in pigs, monkeys and humans. To our knowledge, this was the first report of B. coli infections in sheep in the Qinghai Tibetan Plateau Area of China, or eleswhere around the world. Importantly, the sheep case was rare but raised our concern that B. coli may spread across species and expand its host range.

Phylogenomic analysis of Balantidium ctenopharyngodoni (Ciliophora, Litostomatea) based on single-cell transcriptome sequencing.

In this paper, we present transcriptome data for Balantidium ctenopharyngodoni Chen, 1955 collected from the hindgut of grass carp (Ctenopharyngodon idella). We evaluated sequence quality and de novo assembled a preliminary transcriptome, including 43.3 megabits and 119,141 transcripts. Then we obtained a final transcriptome, including 17.7 megabits and 35,560 transcripts, by removing contaminative and redundant sequences. Phylogenomic analysis based on a supermatrix with 132 genes comprising 53,873 amino acid residues and phylogenetic analysis based on SSU rDNA of 27 species were carried out herein to reveal the evolutionary relationships among six ciliate groups: Colpodea, Oligohymenophorea, Litostomatea, Spirotrichea, Heterotrichea and Protocruziida. The topologies of both phylogenomic and phylogenetic trees are discussed in this paper. In addition, our results suggest that single-cell sequencing is a sound method of obtaining sufficient omics data for phylogenomic analysis, which is a good choice for uncultivable ciliates. The transcriptome data for Balantidium ctenopharyngodoni are the first omics data within the subclass Trichostomatia, and provide a good basis for ciliate phylogenomic analysis, as well as related omics analysis.

Differences between chimpanzee and baboon gastrointestinal parasite communities.

Cross-species infection among humans, chimpanzees (Pan troglodytes) and baboons (Papio spp.) is potentially a significant public health issue in Africa, and of concern in the conservation of P. troglodytes. However, to date, no statistical comparisons have been made between the prevalence, richness and composition of parasite communities in sympatric populations of baboons and P. troglodytes. We compared parasite communities in sympatric P. troglodytes and Papio papio living in a wilderness site, in the Republic of Senegal, West Africa. We asked whether, in the absence of humans, there are significant differences between these hosts in their interactions with gastrointestinal parasites. We tested whether host, location, or time of collection accounted for variation in prevalence, richness and community composition, and compared prevalence across six studies. We concluded that, despite being closely related, there are significant differences between these two hosts with respect to their parasite communities. At our study site, prevalence of Balantidium, Trichuris and Watsonius was higher in P. papio. Papio papio harboured more parasites per host, and we found evidence of a positive association between Trichuris and Balantidium in P. troglodytes but not P. papio.

New insights into the molecular phylogeny of Balantidium (Ciliophora, Vetibuliferida) based on the analysis of new sequences of species from fish hosts.

We obtained sequences of the small subunit ribosomal RNA (rRNA) for two new isolates of Balantidium from fishes, Balantidium polyvacuolum and Balantidium ctenopharingodoni. This is the first introduction of molecular data of Balantidium species from fish hosts in the phylogenetic analyses of the ciliate subclass Trichostomatia. Despite the fact that these species share morphological characteristics common to other species of Balantidium, the phylogenetic analysis of their sequences has shown that they are to be placed in a different branch closely related to the so-called Australian clade. Thus, our results indicate that the genus Balantidium is polyphyletic and possibly should be represented by two different genera; however, the analysis of more species from other poikilothermic hosts (amphibians, reptiles) should be made before a revised taxonomical proposal could be made.

Reisolation and redescription of Balantidium duodeni Stein, 1867 (Litostomatea, Trichostomatia).

In this work, we present reisolation and redescription of Balantidium duodeni Stein, 1867 from the European common brown frog Rana temporaria Linnaeus, 1758 using light and electron microscopy. This species has a unique morphological feature--its cells are flattened along the dorsoventral axis. Because of its unique morphology and localization (duodenum) in the gastrointestinal tract of the host, it has been proposed to recognize B. duodeni as a member of separate genus, Balantidiopsis Penard, 1922. Molecular phylogenetic analysis demonstrates it to be close to the type species Balantidium entozoon (Ehrenberg, 1838). We argue that its placement into separate genus is not substantiated. We also propose to reinstate the genus Balantioides Alexeieff, 1931 with the type species Paramecium coli (Malmstein, 1857). The recently proposed generic name for this taxon, Neobalantidium Pomajbíková et al., 2013, is a junior synonym of the previously recognized name.

Ultrastructural study of Balantidium ctenopharyngodoni Chen, 1955 (Class: Litostomatea) from China with an emphasis on its vestibulum.

A detailed description of the fine structure of Balantidium ctenopharyngodoni Chen, 1955 with an emphasis on its vestibulum is given in the present paper. As to the vestibular kinetids, special attention is paid to the characters of T1, T2 microtubules and nematodesmata. Serving as the major skeleton to the vestibular cortex, the T1, T2 and Pc microtubules are described herein and their support function is also discussed. Moreover, the well-developed nematodesmata of the vestibular kinetids that form a large basket-like complex are described in detail.

Balantidium honghuensis n. sp. (Ciliophora: Trichostomatidae) from the rectum of Rana nigromaculata and R. limnocharis from Honghu Lake, China.

A new trichostome ciliate, Balantidium honghuensis n. sp., was isolated from the recta of Rana nigromaculata and R. limnocharis during parasite surveys in Honghu Lake, Hubei Province, central China in summer of 2010 and 2011. Its detailed morphometric characters based on LM and SEM studies were described herein. The organism is oval in shape and thickly ciliated. The vestibulum is "V" shaped and occupies about 1/3 to 2/5 of the body length. The vestibular and nearby regions possess strong peripheral fibers which form a marked axial fiber about the cytopharynx. More than 10 contractile vacuoles are distributed along the periphery of the latter body. Comparisions were made between this new species and B. sinensis Nie, 1935. They were discriminated from each other in terms of general body forms, body size, and vestibulum shapes. Besides, special attention was paid to its high-speed daughter swarmers which we believed to be the infective stage of B. honghuensis. Possible infection routes of anura amphibian balantidia were discussed.

Novel insights into the genetic diversity of Balantidium and Balantidium-like cyst-forming ciliates.

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. coli in non-human primates using two gene markers (SSrDNA and ITS1-5.8SDNA-ITS2). 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 in the analyses as the closest relatives of B. coli, as well as reference sequences of vestibuliferids. The SSrDNA tree showed the same phylogenetic diversification of B. coli at genus level as the tree constructed based on the ITS region. Based on the polymorphism of SSrDNA sequences, the type species of the genus, namely B. entozoon, appeared to be phylogenetically distinct from B. coli. 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. sulcata with high support, suggesting the existence of a new species within this genus. 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.

Gastrointestinal parasites in greater rheas (Rhea americana) and lesser rheas (Rhea pennata) from Argentina.

Few data exist on the parasites of ratites, especially from regions within their natural range. It is only recently that extensive studies on the parasites of ostriches (Struthio camelus) have been published, mainly from European countries where commercial farming has expanded. Two species of ratites are native in South America: the lesser rhea also known as Darwin's rhea (Rhea pennata) and the greater rhea (Rhea americana). Both species are considered near threatened by the IUCN and are included in the CITES' Appendices I and II, respectively. Parasitological studies have conservation implications, as they allow us to assess the risk of transmission of pathogens from farmed ratites to wild populations. In this study 92 faecal samples from greater rheas and 55 faecal samples from lesser rheas from different localities in Argentine were analyzed to determine their gastrointestinal parasites. In greater rheas the protozoa (Balantidium coli-like and Entamoeba spp.) and helminths (Fasciola hepatica and Deletrocephalus spp.). The protozoa had not previously been cited as parasites of greater rheas in South America. Cysts and/or trophozoites of B. coli-like were found in 16.3% of the samples, while the prevalence of the remaining parasites was below 10%. Lesser rheas harbored the protozoa B. coli-like, Entamoeba spp. and Chilomastix spp. as well as F. hepatica and nematode eggs and larvae. B. coli-like cysts were found in 20.0% of the samples, while the prevalence of the other parasites remained below 5%. Some of them had not been cited as infecting lesser rheas yet.

Balantidium honghuensis n. sp. (Ciliophora: Trichostomatidae) from the Rectum of Rana nigromaculata and R. limnocharis from Honghu Lake, China

A new trichostome ciliate, Balantidium honghuensis n. sp., was isolated from the recta of Rana nigromaculata and R. limnocharis during parasite surveys in Honghu Lake, Hubei Province, central China in summer of 2010 and 2011. Its detailed morphometric characters based on LM and SEM studies were described herein. The organism is oval in shape and thickly ciliated. The vestibulum is "V" shaped and occupies about 1/3 to 2/5 of the body length. The vestibular and nearby regions possess strong peripheral fibers which form a marked axial fiber about the cytopharynx. More than 10 contractile vacuoles are distributed along the periphery of the latter body. Comparisions were made between this new species and B. sinensis Nie, 1935. They were discriminated from each other in terms of general body forms, body size, and vestibulum shapes. Besides, special attention was paid to its high-speed daughter swarmers which we believed to be the infective stage of B. honghuensis. Possible infection routes of anura amphibian balantidia were discussed.

[Population characterization of Balantidium coli from pigs using ITS1-5.8S rRNA-ITS2 sequence].

OBJECTIVE: To determine the species and genotypes of Balantidium isolated from pigs in Henan province, China. METHODS: Scatoscopy and the modified DMEM media were used to isolate trophozoites of Balantidium from pig feces. The ITS1-5.8S rRNA-ITS2-based molecular marker method, Acridine orange staining ( AO) and microscopic observation were used to determine the population characteristics among different isolates of B. coli from various pigs farms. RESULTS: We isolated 15 isolates from the pigs at diagnosis in the Animal Hospital of Henan University of Science and Technology from the pig farms of the 8 counties or cities of the west of Henan province in total, and all of them belong to the same species B. coli. MJ-2 and SX-1 isolates were genotype A of B. coli, and the remaining 13 isolates were genotype B. Trophozoites of MJ-2 and SX-1 were bigger, moved more slowly and lower density in feces and in vitro culture than other 13 isolates, while structures of their nuclei were not different. CONCLUSION: Both genotype A and B of B. coli are present in the pig farms of the west of Henan province, China, and genotype B is the determinant population in pigs farms. These findings could provide an important implication for the effective control of balantidiosis of human and other hosts.

Ultrastructural and molecular characterization of Balantidium coli isolated in the Philippines.

Balantidium coli is a ciliated protozoon inhabiting the colon of swine, rodents, horses, nonhuman primates and humans. In association with disease triggered by other infectious agents, B. coli may become a pathogenic opportunist. This study describes the isolation, cultivation, morphological as well as molecular characterization of B. coli isolated from the large intestine of a pig in the Philippines. Based on scanning and transmission electron microscopy, this protozoon presents a dense ciliation in the oral structure and somatic cilia that are arranged in a more transverse field. Oral and somatic monokinetids were identified in the cortex of the organism. The presence of heterokaryotic nuclear condition is evident, and the cell body of the ciliate shows numerous mucocysts, several food vacuoles, mitochondria, endoplasmic reticulum, and contractile vacuoles. Polymerase chain reaction and phylogenetic analysis based on the small subunit ribosomal RNA gene were performed in order to compare our isolate with other previously reported B. coli isolates. The full-length sequence of the SSU rRNA gene of the isolate showed 99% similarity to other B. coli isolates reported in the GenBank. Phylogenetic analysis revealed that the isolate clustered with previously reported B. coli isolates from gorillas, pig, and ostrich. To date, no studies on the ultrastructure and phylogeny of B. coli isolated in the Philippines have been reported. Results from this study may serve as a baseline data for further ultrastructural and phylogenetic studies on this organism. This study also suggests that morphological characteristics along with molecular identification are essential for validating and identifying species of Balantidium.

A re-description of the ciliate genus and type species, Balantidium entozoon.

Members of the ciliate genus Balantidium possess a specialized "Villeneuve-Brachon's" field of somatic cilia to the right of the vestibule, or in a dextroral location. Specimens of the type species were collected in Italy and fixed for study by light microscopy, and scanning and transmission electron microscopy. The presence of the field in the type species and several other species of the genus indicates a need to re-describe the genus by including details of the ultrastructure of that field. Scanning electron microscopy shows that the field consists of one row of relatively short cilia of uniform length flanked on each side by 2-3 rows, or more, of very short cilia. Their kinetids have typical litostome structure in transmission electron micrographs. We speculate on a possible function for the Villeneuve-Brachon's field and also present morphometric data on the type species. The base sequence of the small subunit ribosomal RNA gene of Balantidium entozoon has been determined and found to differ by 5% from that of B. coli. Based on the location and ultrastructure, organelles found around the somatic kinetosomes and within inter-kinetal ridges of B. entozoon were identified as hydrogenosomes.

First report of two Balantidium species from the Chinese giant salamander, Andrias davidianus: Balantidium sinensis Nie 1935 and Balantidium andianusis n. sp.

Two ciliated protozoa, Balantidium sinensis Nie 1935 and Balantidium andianusi n. sp., were isolated from the feces of a wild Chinese giant salamander (Andrias davidianus) captured from the mountainous area of Shiyan, Hubei Province, Central China in October 2006. It is the first report of Balantidium species inhabiting Cryptobranchoidea amphibians. The occurrence of B. sinensis in A. davidianus should be a new record because the type specimens were first discovered and named by Nie in 1935 from Rana nigromaculata and Rana plancyi. For the lack of enough descriptions of taxonomic features in the previous report, it was re-described in detail and compared with Nie's type specimens and B. giganteum to complete the morphological descriptions in the present work. B. andianusi n. sp. was considered to be a new species based on its unique morphological characteristics, especially the high length/width ratio of the vestibulum (8:1). Comparisons were also made among Balantidium species that were found from urodele amphibians.

Amebae and ciliated protozoa as causal agents of waterborne zoonotic disease.

The roles free-living amebae and the parasitic protozoa Entamoeba histolytica and Balantidium coli play as agents of waterborne zoonotic diseases are examined. The free-living soil and water amebae Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris are recognized etiologic agents of mostly fatal amebic encephalitides in humans and other animals, with immunocompromised and immunocompetent hosts among the victims. Acanthamoeba spp. are also agents of amebic keratitis. Infection is through the respiratory tract, breaks in the skin, or by uptake of water into the nostrils, with spread to the central nervous system. E. histolytica and B. coli are parasitic protozoa that cause amebic dysentery and balantidiasis, respectively. Both intestinal infections are spread via a fecal-oral route, with cysts as the infective stage. Although the amebic encephalitides can be acquired by contact with water, they are not, strictly speaking, waterborne diseases and are not transmitted to humans from animals. Non-human primates and swine are reservoirs for E. histolytica and B. coli, and the diseases they cause are acquired from cysts, usually in sewage-contaminated water. Amebic dysentery and balantidiasis are examples of zoonotic waterborne infections, though human-to-human transmission can occur. The epidemiology of the diseases is examined, as are diagnostic procedures, anti-microbial interventions, and the influence of globalization, climate change, and technological advances on their spread.

New species of Balantidium and Paracichlidotherus (Ciliophora) inhabiting the intestines of four surgeonfish species from the Tuvalu Islands, Pacific Ocean.

Four species of adult herbivorous surgeonfishes (Family Acanthuridae) were collected from the remote South-Pacific island system of Tuvalu. Their intestinal contents were examined, and of four populations of ciliated protists, two new species were discovered and are described. Ciliates were examined after protargol staining and, in some cases, scanning electron microscopy. Members of each population were examined and 10 characters measured for the balantidia, and 13 for the paracichlidotherids. A new Balantidium is described which has an unusually large dextr-oral field of cilia. A new species of Paracichlidotherus was discovered which has a macronucleus significantly smaller and well anteriad the cytoplasmic portion of the oral polykinetids relative to the type species.