Interactions between Balantidium ctenopharyngodoni and microbiota reveal its low pathogenicity in the hindgut of grass carp.

BACKGROUND: Hosts, parasites, and microbiota interact with each other, forming a complex ecosystem. Alterations to the microbial structure have been observed in various enteric parasitic infections (e.g. parasitic protists and helminths). Interestingly, some parasites are associated with healthy gut microbiota linked to the intestinal eubiosis state. So the changes in bacteria and metabolites induced by parasite infection may offer benefits to the host, including protection from other parasitesand promotion of intestinal health. The only ciliate known to inhabit the hindgut of grass carp, Balantidium ctenopharyngodoni, does not cause obvious damage to the intestinal mucosa. To date, its impact on intestinal microbiota composition remains unknown. In this study, we investigated the microbial composition in the hindgut of grass carp infected with B. ctenopharyngodoni, as well as the changes of metabolites in intestinal contents resulting from infection. RESULTS: Colonization by B. ctenopharyngodoni was associated with an increase in bacterial diversity, a higher relative abundance of Clostridium, and a lower abundance of Enterobacteriaceae. The family Aeromonadaceae and the genus Citrobacter had significantly lower relative abundance in infected fish. Additionally, grass carp infected with B. ctenopharyngodoni exhibited a significant increase in creatine content in the hindgut. This suggested that the presence of B. ctenopharyngodoni may improve intestinal health through changes in microbiota and metabolites. CONCLUSIONS: We found that grass carp infected with B. ctenopharyngodoni exhibit a healthy microbiota with an increased bacterial diversity. The results suggested that B. ctenopharyngodoni reshaped the composition of hindgut microbiota similarly to other protists with low pathogenicity. The shifts in the microbiota and metabolites during the colonization and proliferation of B. ctenopharyngodoni indicated that it may provide positive effects in the hindgut of grass carp.

Positive clinical experience with paromomycin sulfate in treatment of Balantioides coli (=Balantidium coli) natural infection in zoo-kept Mandrill monkeys (Mandrillus sphinx) and Western lowland gorillas (Gorilla gorilla gorilla).

Balantioides coli (=Balantidium coli), a large ciliated protozoan, is reported in multiple free-ranging and captive primate species, often in association with a clinical presentation that requires medical intervention. This report describes the clinical effectiveness of paromomycin sulfate against B.coli in zoo-kept mandrill monkeys (Mandrillus sphinx, at orally doses of 8-31 mg/kg, once daily (SID) for 7 days) and gorillas (Gorilla gorilla gorilla, at orally doses of 1.4-3.1 mg/kg, SID for 5 days).

The energy metabolism of Balantidium polyvacuolum inhabiting the hindgut of Xenocypris davidi.

Anaerobic parasitic ciliates are a specialized group of ciliates that are adapted to anoxic and oxygen-depleted habitats. Among them, Balantidium polyvacuolum, which inhabits the hindgut of Xenocyprinae fishes, has received very limited scientific attention, so the molecular mechanism of its adaptation to the digestive tract microenvironment is still unclear. In this study, transmission electron microscopy (TEM) and single-cell transcriptome analysis were used to uncover the metabolism of B. polyvacuolum. Starch granules, endosymbiotic bacteria, and multiple specialized mitochondrion-related organelles (MROs) of various shapes were observed. The MROs may have completely lost the electron transport chain (ETC) complexes I, III, IV, and V and only retained succinate dehydrogenase subunit A (SDHA) of complex II. The tricarboxylic acid (TCA) cycle was also incomplete. It can be inferred that the hypoxic intestinal environment has led to the specialization of the mitochondria in B. polyvacuolum. Moreover, carbohydrate-active enzymes (CAZymes), including carbohydrate esterases, enzymes with a carbohydrate-binding module, glycoside hydrolases, and glycosyltransferases, were identified, which may constitute evidence that B. polyvacuolum is able to digest carbohydrates and starch. These findings can improve our knowledge of the energy metabolism and adaptive mechanisms of B. polyvacuolum.

Identification of Intracellular Bacteria in the Ciliate Balantidium ctenopharyngodoni (Ciliophora, Litostomatea).

The ciliate Balantidium ctenopharyngodoni is the most prominent protist in the guts of grass carp, where it mainly inhabits the creamy luminal contents of the hindgut. Ciliates are generally colonized by microorganisms via phagotrophic feeding. In order to study the intracellular bacteria in this ciliate, we have successfully established it in in vitro culture. Herein, we investigated and compared the bacterial community structures of cultured and freshly collected B. ctenopharyngodoni. The results showed that these two groups exhibited different bacterial communities. The most abundant bacterial family in freshly collected samples was Enterobacteriaceae, while in cultured samples it was Fusobacteriaceae. In addition, a key intracellular bacterium, Cetobacterium somerae, was identified in the cytoplasm of cultured ciliates using fluorescence in situ hybridization (FISH). This study shows that ciliates can retain the intracellular bacteria acquired in the natural habitat for quite a long time, but the bacterial community structure of ciliates eventually changes after a long period of cultivation.

Dysentery Caused by Balantidium coli in China.

Balantidium coli human infection predominantly occurs in tropical and subtropical regions in the world. Human case is extremely rare in China. This report details a case of B. coli infection in a 68-year-old man in China, who presented with history of abdominal pain, tenesmus, diarrhea with blood and was diagnosed as B. coli-caused dysentery. Our case indicates possible occurrence of Balantidium coli-related disease in cooler climates. This case is presented not only because of its rarity but also for future references.

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.

Occurrence of anthropozoonotic parasitic infections and faecal microbes in free-ranging sperm whales (Physeter macrocephalus) from the Mediterranean Sea.

Sperm whales (Physeter macrocephalus) are the largest toothed whales and only living member of family Physeteridae. Present survey represents first report on cultivable faecal microbes and gastrointestinal helminths and protozoans infecting free-ranging sperm whales inhabiting Mediterranean Sea waters surrounding Balearic Archipelago, Spain. Twenty-five individual sperm whale scat samples, including one calf, were collected without disturbance of animals during the summer of 2016. Parasitological diagnostic methods, such as sodium acetate acetic formalin (SAF) method, carbol fuchsin-stained faecal smears, Giardia/Cryptosporidium coproantigen ELISAs and an Anisakis-specific PCR were applied for further identification. Five bacterial genera, i.e. Acinetobacter, Clostridium, Enterococcus, Staphylococcus and Streptococcus, and one fungus namely Cladosporium were identified. Parasitological infections included seven different parasite species with some of them bearing anthropozoonotic potential. Thus, four of these parasites were zoonotic, i.e. Anisakis, Balantidium, Diphyllobothriidae gen. sp. and Giardia. Additionally, Zalophotrema curilensis eggs, spirurid-like eggs and Cystoisospora-like oocysts were identified. Molecular characterization identified Anisakis physeteris as the species infecting these whales. This survey provides first records on occurrence of two zoonotic enteropathogenic protozoan parasites (Giardia and Balantidium) and of facultative pathogenic bacteria (Clostridium and Enterococcus) in sperm whales. Presented data should be considered as a baseline study for future monitoring surveys on anthropozoonotic pathogens affecting free-living sperm whale populations and enhance investigations on possible impact on public health as well as on isolated Mediterranean sperm whale subpopulation.

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.

A Comparative Study of Four Methods for the Detection of Nematode Eggs and Large Protozoan Cysts in Mandrill Faecal Material.

Coproscopical methods like sedimentation and flotation techniques are widely used in the field for studying simian gastrointestinal parasites. Four parasites of known zoonotic potential were studied in a free-ranging, non-provisioned population of mandrills (Mandrillus sphinx): 2 nematodes (Necatoramericanus/Oesophagostomum sp. complex and Strongyloides sp.) and 2 protozoan species (Balantidium coli and Entamoeba coli). Different coproscopical techniques are available but they are rarely compared to evaluate their efficiency to retrieve parasites. In this study 4 different field-friendly methods were compared. A sedimentation method and 3 different McMaster methods (using sugar, salt, and zinc sulphate solutions) were performed on 47 faecal samples collected from different individuals of both sexes and all ages. First, we show that McMaster flotation methods are appropriate to detect and thus quantify large protozoan cysts. Second, zinc sulphate McMaster flotation allows the retrieval of a higher number of parasite taxa compared to the other 3 methods. This method further shows the highest probability to detect each of the studied parasite taxa. Altogether our results show that zinc sulphate McMaster flotation appears to be the best technique to use when studying nematodes and large protozoa.

Endoparasite survey of free-swimming baleen whales (Balaenoptera musculus, B. physalus, B. borealis) and sperm whales (Physeter macrocephalus) using non/minimally invasive methods.

A number of parasitic diseases have gained importance as neozoan opportunistic infections in the marine environment. Here, we report on the gastrointestinal endoparasite fauna of three baleen whale species and one toothed whale: blue (Balaenoptera musculus), fin (Balaenoptera physalus), and sei whales (Balaenoptera borealis) and sperm whales (Physeter macrocephalus) from the Azores Islands, Portugal. In total, 17 individual whale fecal samples [n = 10 (B. physalus); n = 4 (P. macrocephalus); n = 2 (B. musculus); n = 1 (B. borealis)] were collected from free-swimming animals as part of ongoing studies on behavioral ecology. Furthermore, skin biopsies were collected from sperm whales (n = 5) using minimally invasive biopsy darting and tested for the presence of Toxoplasma gondii, Neospora caninum, and Besnoitia besnoiti DNA via PCR. Overall, more than ten taxa were detected in whale fecal samples. Within protozoan parasites, Entamoeba spp. occurred most frequently (64.7%), followed by Giardia spp. (17.6%) and Balantidium spp. (5.9%). The most prevalent metazoan parasites were Ascaridida indet. spp. (41.2%), followed by trematodes (17.7%), acanthocephalan spp., strongyles (11.8%), Diphyllobotrium spp. (5.9%), and spirurids (5.9%). Helminths were mainly found in sperm whales, while enteric protozoan parasites were exclusively detected in baleen whales, which might be related to dietary differences. No T. gondii, N. caninum, or B. besnoiti DNA was detected in any skin sample. This is the first record on Giardia and Balantidium infections in large baleen whales.

Molecular genetic investigations on Balantidium ctenopharyngodoni Chen, 1955, a parasite of the grass carp (Ctenopharyngodon idella).

Balantidium ctenopharyngodoni is a common ciliate in Hungary, infecting the hindgut of grass carp (Ctenopharyngodon idella), a cyprinid fish of Chinese origin. Although data have already been presented on its occasional pathogenic effect on the endothelium of the host, generally it is a harmless inhabitant of the gut. Phylogenetic analysis of the 18S rDNA and ITS fragments of this protozoan proved that it is in the closest phylogenetic relationship with endocommensalist and symbiont ciliates of mammals feeding on large volumes of green forage, in a similar way as Balantidium spp. known from algae-eating marine fishes.

Balantidium coli and other gastrointestinal parasites in captives non-human primates of the Rio de Janeiro, Brazil.

BACKGROUND: Parasites are agents of diarrhea in captive non-human primates (NHP). To broaden information about those etiological agents in captive animals in Brazil, gastrointestinal parasites in captive NHP were researched in nurturing Centers of Rio de Janeiro State. METHODS: Fecal samples were collected from primates, of which 960 came from the Research Center (Cecal/Fiocruz) and 115 from the Primate Center (CPRJ/Inea). The study involved species of the New World (NW) primates and of the Old World (OW). RESULTS: The estimated prevalence was 56.7%, of which 91.3% presented protozoans and 7.4% presented helminths. Statistical difference between the nurturing centers occurred in the overall value of parasitosis and in the isolated frequency of Balantidium coli and Entamoeba sp., especially in the samples of OW primates living in Cecal. CONCLUSIONS: These results demonstrated the need for implements of sanitation programs in the sites for captive primates nurturing.

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.

Prevalence of Balantidium coli infection in sows in Hunan province, subtropical China.

Balantidium coli is a cosmopolitan parasitic-opportunistic pathogen that can be found throughout the world. However, little information is available about prevalence of B. coli in pigs in China. In the present study, the prevalence of B. coli in pigs was investigated in Hunan province, subtropical China, between January 2012 and August 2014. A total of 3925 diarrheic fecal samples from nine representative administrative regions in Hunan province, subtropical China, were examined for the presence of B. coli cysts and/or trophozoites using microscopy after sedimentation with water. The overall prevalence of B. coli in pigs was 36.9 % (1450/3925). The present survey revealed high circulation of B. coli in pigs in Hunan province, subtropical China, which poses potential threats to human health. The results of the present investigation have important implications for the control of B. coli infections in pigs in Hunan province, subtropical China. To our knowledge, this is the first comprehensive report of B. coli prevalence in sows in Hunan province, subtropical China.

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.

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.

Apparent mortality as a result of an elevated parasite infection in Presbytis rubicunda.

We provide the first parasite assessment of wild red langurs (Presbytis rubicunda). Seven helminth taxa and 4 protozoa were recovered from faecal samples. We report the details of the death of an adult female red langur with potential association of an elevated parasite infection. There is a paucity of published literature on red langurs and nothing is known of the factors affecting interspecific parasite transmission and disease spread between other primate species. Parasites may be substantial determinants of host health and may present a significant influence on the survival and reproduction of individuals and the subsequent fecundity of populations. Further research into the connection of parasite burden with individual deaths and potential population declines is recommended for monitoring population health.

[The study of Balantidium Helenae Bezzenberger, 1904 (Litostomatea, Trichostomatia) inhabiting the intestines of the frog Rana temporaria Linnaeus, 1758, in optical and electron microscopes].

The structure of the ciliate Balantidium helenae Bezzenberger, 1904 was investigated with the use of optical and electron microscopy. It was demonstrated that, together with the basic dimensional characteristics, main distinguishing features of the studied species include the following characters: the presence of a pointed prominence on the posterior margin of the cell; the presence of elongated cilia on the left margin of the vestibulum; and different shape and size of cortical ridges. Two morphologically different types of electron-dense bodies (supposedly hydrogenosoms) and also endobiotic bacteria were also found in the cytoplasm of B. helenae.

Genome assembly of a symbiotic balantidia (Balantidium ctenopharyngodoni) in fish hindgut.

Balantidium ctenopharyngodoni is identified as the sole ciliate species that exclusively resides within the hindgut of grass carp with high prevalence and intensity. In this study, the successful cultivation of B. ctenopharyngodoni enabled us to collect enough cells for genome sequencing. Consequently, we acquired a high-quality genome assembly spanning 68.66 Mb, encompassing a total of 22,334 nanochromosomes. Furthermore, we predicted 29,348 protein-coding genes, and 95.5% of them was supported by the RNA-seq data. The trend of GC content in the subtelomeric regions of single-gene chromosomes was similar to other ciliates containing nanochromosomes. A large number of genes encoding carbohydrate-binding modules with affinities for starch and peptidoglycans was identified. The identification of mitochondrion-related organelles (MROs) within genome indicates its well-suited adaptation to the anaerobic conditions in the hindgut environment. In summary, our results will offer resources for understanding the genetic basis and molecular adaptations of balantidia to hindgut of herbivorous fish.

Isolated Balantidium coli in a 1.5-Year-Old Boy: A Case Report.

PURPOSE: Balantidium coli, is a protozoa that can infect humans. Balantidium coli infection is mostly asymptomatic, but can cause of diarrhea, abdominal pain and sometimes perforated colon. There are only few previous reports regarding the infant balantidiasis and there is a little information about its clinical signs and treatment. In Islamic countries pig farming and consumption is prohibited and reservoir host is unknown. METHODS: 1.5 years old boy showed sever anorexia and diarrhea. The affected infant was depressed. The evaluation of fecal examination revealed a large number of B. coli trophozoites and cysts leaded us to an uncommon presentation of B. coli in the infant. RESULTS: A few reports of human balantidiasis from Iran seems quite extraordinary. Wild boars have been proposed the responsible for transmission of human infection in Iran. CONCLUSION: However, this report as the first report of infant balantidiasis in Iran supports the role of other sources and domestic or wild animals as reservoir hosts for B. coli in Iran.