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.

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.

[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.

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.

Light and scanning electron microscopic study of Balantidium ctenopharyngodoni Chen, 1955 (Class: Litostomatea) from China.

Redescription of Balantidium ctenopharyngodoni "Chen (Acta Hydrobiol Sin 1:123-164, 1955)", collected from the hindgut of grass carp (Ctenopharyngodon idella), especially the segment of 6-10 cm upstream from the anus, from Honghu Lake, Hubei Province, central China in November 2005, is presented in this paper to complete Chen's description at both light and scanning electron microscopic levels. Some revisions were done: the vestibulum is fairly symmetrical, with compactly arranged cilia rather than assembled membrane bordering on the left vestibular side; four contractile vacuoles actually exist in the latter body, three of which surround the posterior portion of the macronucleus, whereas the fourth lies antero-left to it. Somatic monokinetids were compared among the species of genus Balantidium. The cysts were described, and possible infection routes of B. ctenopharyngodoni were also discussed.

Food vacuole contents in the ciliate, Balantidium jocularum (Balantididae), a symbiont in the intestine of the surgeonfish, Naso tonganus (Acanthuridae).

During the past 16 years, the ciliate Balantidium jocularum has been collected from the intestines of many specimens of its fish host, Naso tonganus, all collected from the Great Barrier Reef near Lizard Island, Australia. Ciliates for this study of food consumption were isolated in 1988, 1989, 2003, and 2005. Nineteen specimens of B. jocularum were examined in the transmission electron microscope to determine the contents of both food vacuoles and a putative discharging cytoproct vacuole. Food vacuoles contained rod-shaped bacteria, tightly coiled spirilliform bacteria, and one or more euglenid flagellates. In several balantidia of somewhat different form than the type species of B. jocularum, the large bacterium, Epulopiscium fishelsoni, was observed in light microscope protargol preparations. Some putative phagolysosomes retained spirilliform bacteria that were apparently intact, and others contained partially digested flagellates. Food in a single discharging cytoproct vacuole consisted of normal appearing spirilliform bacteria, some other bacteria, and no flagellates. The results argue for non-selective ingestion of food and selective digestion; hence, somewhat inefficient food processing.

An electron microscopic study of the phosphatases in the ciliate Balantidium coli.

The localisation and activity of D glucose-6-phosphatase (G-6-Pase) and alkaline phosphatase (AlP) in the trophozoites of Balantidium coli isolated from pig intestine content were investigated using ultrastructural and cytochemical methods. The activity of G-6-Pase was demonstrated on the membranes of the endoplasmic reticulum, particularly in the cortical part of the trophozoites. In addition, the product of the reaction to G-6-Pase was concentrated in the vesicular structures, which were distributed along the reticular membranes. These structures were described as vesicles similar to glycosomes, containing enzymes of glycogenolysis. It is very likely that hydrolases in B. coli are formed on the rough reticular membranes without the involvement of cisterns of the Golgi complex. The ultrastructural deposits of the reaction to G-6-Pase and AlP in the trophozoites of B. coli described here indicate that some membranes of the rough endoplasmic reticulum and small vacuoles with a strong reaction to these enzymes can play a similar role to the Golgi complex.

The formation of primary and secondary lysosomes in Balantidium coli, Ciliata.

Trophozoites, vegetative forms of Balantidum coli isolated from pigs affected by acute and asymptomatic balantidiasis were studied. Lysosomes and food vacuoles were revealed by cytochemical detection of lysosomal marker, acid phosphatase. The cytoplasm of all the B. coli trophozoites examined was found to contain numerous structures which differed widely in shape, size and location in the cells. One of them was located among the rough endoplasmic reticulum membranes and another one in the vicinity of endosomes. Those structures were regarded as the primary lysosomes. The two types of vesicular structures most probably represent two stages of the primary lysosome formation. Trophozoites were also found to contain secondary lysosomes which are formed by fusion of several primary lysosomes with phagosomes. The ultrathin sections of B. coli trohozoites showed the presence of two types of phagosomes. They were divided, based on their contents, into auto- and heterophagosomes.

Cytochemical identification of mucocysts in Balantidium coli trophozoites.

The mucocyst ultrastructure in B. coli has not been described so far. As demonstrated in this work, cytoenzymatic assays on B. coli with the use of a reaction-detecting membrane-coupled hydrolase, i.e., ATP-ase, permitted identification of the mucocysts in the ciliate studied. The shape, size, and location of mucocysts in B. coli trophozoites were found to correspond to descriptions of these structures in other ciliates. The mucocysts were more numerous in B. coli trophozoites isolated from the symptomatic balantidiosis-affected pigs (Group I), and the product of reaction to ATP-ase was more copious than in Group II trophozoites. However, not all the bubble-like structures with similar morphological features reacted positively to the enzyme. The discrepancy was explained by the cytoenzymatic reaction to Beta-GR. The reaction product was visible in the vesicular structures, situated above the plasmolemma, although some of them contained no reaction product. Thus the presence of two types of secretory structure can be inferred: the mucocysts, with ATP-ase in their membranes, and other extrusomes containing active Beta-GR.

Ultrastructural and cytochemical identification of peroxisomes in Balantidium coli, Ciliophora.

Peroxisomes of the trophozoites of Balantidium coli isolated from pig intestine content were investigated, using ultrastructural and cytochemical techniques. The peroxisomes of B. coli trophozoites from pigs with subclinical balantidiasis are less then 0.8 mm in diameter whereas those from pigs with acute balantidiasis are greater than 0.8 micron in diameter. In all the trophozoites peroxisomes are round, oval or dumb-bell shaped. Catalase as an indicative enzyme was detected by cytochemical techniques in B. coli peroxisomes.