The complete life cycle of the unusual apostome Hyalophysa clampi (Ciliophora,Apostomatida), a symbiont of crayfish in Alabama (USA)

Hyalophysa clampi Browning and Landers, 2012 was reexamined to determine all stages in the life cycle of this symbiotic ciliate. The cell feeds as a normal exuviotroph within the exoskeleton of its molted crayfish host but does not encyst following the trophont stage. Trophonts transform into swimming tomont stages, which divide by palintomy over successive divisions, splitting to two cells, separating, and repeating. The divisions cease when the daughter cells attain the size of the infestive tomite stage, which attaches to a new crayfish. This unique life cycle is most similar to the European hermit crab symbiont Polyspira delagei, which forms chains of daughter cells during division. Scanning electron microscopy confirmed the unusual presence of two contractile vacuoles in H. clampi, unique among the Apostomatida, and provided ultrastructural details to better understand light microscopy silver staining. The genus diagnosis for Hyalophysa is modified herein to accommodate this new life cycle.     

Exuviotrophic apostome ciliates from crustaceans of St. Andrew Bay, Florida, and a description of Gymnodinioides kozloffi n. sp

Gymnodinioides kozloffi n. sp. is described from the eelgrass broken-back shrimp Hippolyte zostericola. The species is distinct from others in the apostome genus Gymnodinioides in that the trophont ciliature has a small group of kinetosomes located to the right of Kinety 9a, and Kinety 1 and 2 are divided. Other apostome morphologies are described from many decapod crustaceans from St. Andrew Bay, Florida, including Gymnodinioides inkystans, Hyalophysa chattoni, and variants of both H. chattoni and G. kozloffi. All of these apostome ciliates are exuviotrophic, found feeding on exuvial fluid within the exoskeleton of the host after ecdysis. The hosts surveyed for this study are the following: Callinectes sapidus, Eurypanopeus depressus, Hippolyte zostericola, Farfantepenaeus spp., Palaemonetes intermedius, Palaemon floridanus, Portunus spp., Tozeuma carolinense, and Sicyonia laevigata, which revealed a number of new host-apostome records.     

Hyalophysa lynni n. sp. (Ciliophora,Apostomatida), a new pathogenic ciliate and causative agent of shrimp black gill in penaeid shrimp

The parasitic ciliate causing shrimp black gill (sBG) infections in penaeid shrimp has been identified. The sBG ciliate has a unique life cycle that includes an encysted divisional stage on the host’s gills. The ciliature of the encysted trophont stage has been determined and is quite similar to that of the closely related apostomes Hyalophysa bradburyae and H. chattoni. Hyalophysa bradburyae is a commensal ciliate associated with freshwater caridean shrimp and crayfish, while H. chattoni is a common commensal found on North American marine decapods. Based on 18S rRNA gene sequence comparisons, the sBG ciliate is more closely related to the marine species H. chattoni than to the freshwater species H. bradburyae. The unique life cycle, morphology, 18S rRNA gene sequence, hosts, location, and pathology of the sBG ciliate distinguish this organism as a new species, Hyalophysa lynni n. sp.     

Taxonomic characterization of two marine peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp. (Ciliophora: Peritrichia), from North China

The morphology, infraciliature, and silverline system of two new peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp., have been investigated based on both living and silver-impregnated specimens. Partial sequence of 18S-ITS1-5.8S rDNA of Z. pararbuscula is also determined in order to compare it with the closely related congener, Zoothamnium arbuscula. Zoothamnium pararbuscula can be distinguished from its close form Z. arbuscula by the different habitats, the appearance of the main stalk, the position of the contractile vacuole, and the information derived from 18S-ITS1-5.8S rDNA sequence analysis. Pseudovorticella clampi n. sp. is distinguished from its congeners by its body shape and size, pellicle granules, habitat, and number of transverse silverlines.     

The fine structure of the hypertrophont of the parasitic apostome Synophrya (Ciliophora,Apostomatida)

The hypertrophont stage of the parasitic apostome ciliate Synophrya was studied by light microscopy and transmission electron microscopy. This invasive stage of the ciliate was found within the gill lamellae and gill raphes of the longspine swimming crab Portunus spinicarpus. The ciliates elicited a melanized host reaction that walled off the parasite from the host. Additionally the ciliate produced a cyst wall of ～0.16-2.0 μm in thickness that further isolated the parasite. A mouth was not observed, as the internal stage of this ciliate takes in material via endocytosis across the entire surface. The outer surface was irregular, with folds, membrane pillows, and vesicles connected to the outer membrane. The hypertrophont had a sparce ciliature, with well developed kinetodesmal fibers connecting the kinetosomes. Within the cytoplasm the cell had numerous vacuoles, lipid droplets, and large plaquettes of material. The massive reticulate macronucleus had globular and elongated chromatin bodies, and was the most distinctive organelle within the cell.     

Morphologic and molecular characterization of Apertospathula pilata n. sp., a novel freshwater spathidiid (Ciliophora,Litostomatea) from Idaho,USA

Order Spathidiida Foissner and Foissner, 1988 comprises a large group of morphologically diverse, primarily predatory, free living ciliates, the phylogeny of which has remained stubbornly unresolved. Families Arcuospathidiidae and Apertospathulidae are two morphologically similar groups established on the basis of differences in the morphology of the oral bulge and circumoral kinety. While Arcuospathidiidae is non-monophyletic in 18S rRNA gene analyses, the Apertospathulidae has been represented by only a single Apertospathula sequence in public databases. In this report, a novel freshwater species, Apertospathula pilata n. sp. is described on the basis of living observation, silver impregnation, and scanning electron microscopy. The phylogeny of the new species is assessed based on the rRNA cistron. The main features distinguishing A. pilata n. sp. from all congeners are: the oral bulge extrusomes (filiform, up to 25 µm long), the combination of body size (130–193 µm) and shape (spatulate), the extensive oral bulge length (41% of the cell length after protargol impregnation), and multiple micronuclei (one to five, two on average). The monophyly of Apertospathulidae Foissner, Xu and Kreutz, 2005 is rejected.     

Gymnodinioides pacifica, n. sp., an exuviotrophic ciliated protozoan (Ciliophora, Apostomatida) from euphausiids of the Northeastern Pacific

A new species of apostome ciliate, Gymnodinioides pacifica n. sp. from the euphausiid Euphausia pacifica is described. The ciliated protozoan encysts on the setae of the appendages, telson and antennae. It excysts and enters the exoskeleton of the host after moulting, where it feeds on exuvial fluid. The phoront and trophont stages of this ciliate are described. The ciliature of the trophont has the following characteristics that distinguish it from the other species of Gymnodinioides: (1) a short kinety 9a (ciliary row 9a or K9a), approximately half of the length of falciform field 8, that extends from the anterior end of the cell posteriorly to the level of K1, (2) a straight K5a, perpendicular to K5b, K6, and K7, and (3) two distinct bands of kineties, K9b-K5b and K4-K1. In addition to those on the host E. pacifica, phoront cysts of similar shape were observed on Thysanoessa spinifera, T. gregaria, T. inspinata, T. longipes, and Nematoscelis difficilis. High prevalence rates were recorded (83%) for these apostome cysts on E. pacifica and T. spinifera during summer collections from the Oregon and Washington coasts. Additionally, we report other apostome ciliate symbionts of euphausiids, including Phtorophrya sp., which preys on apostomes in the genus Gymnodinioides.     

Studies of the Phoront of Hyalophysa chattoni (Ciliophora,Apostomatida) Encysted on Grass Shrimp1

ABSTRACT. The apostomatous ciliate Hyalophysa chattoni, an ectosymbiont of the grass shrimp Palaemonetes pugio, encysts and dedifferentiates within 48 h from the migratory tomite to a phoretic stage devoid of complex ciliary fields. The presettlement crawling and pivoting of the tomite may play a role in its initial attachment to the shrimp. Metamorphosis of exuviotrophic apostomes has been previously observed to take place immediately prior to host ecdysis. The study has found that Hyalophysa's metamorphosis to the feeding stage on grass shrimp is initiated by a cue from the premolt host and begins during earlier stages of the molt cycle (D₀ and D₁). Due to the long premolt stage of the host's diecdysic molt cycle, metamorphosis is initiated well before ecdysis (over six days). Hyalophysa was able to encyst and metamorphose within 41/4 h when exposed to shrimp in a late premolt stage, indicating that the control of apostome metamorphosis is solely host-dependent.     

Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio

The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations.     

Morphology and infraciliature of two new marine urostylid ciliates: Metaurostylopsis struederkypkeae n. sp. and Thigmokeronopsis stoecki n. sp. (Ciliophora, Hypotrichida) from China

ABSTRACT. The morphology and infraciliature of two new marine urostylid ciliates, Metaurostylopsis struederkypkeae n. sp. and Thigmokeronopsis stoecki n. sp., collected from the coastal waters off Qingdao (Tsingtao), China, have been investigated. Metaurostylopsis struederkypkeae n. sp. is characterized by the slender body shape, small size, rose-reddish cell colour, and having two kinds of pigment-like granules. The larger pigment-like granules are yellow–green or grass-green in colour, oval in shape, and flattened, whereas the smaller ones are wine-reddish. Infraciliature and nuclear apparatus are similar to the well-known Metaurostylopsis marina. Thigmokeronopsis stoecki n. sp. is characterized by its large size with dark brown cell colour and grass-green cortical granules, which are large, blood-cell shaped, and sparsely distributed. The thigmotactic ciliature is conspicuous: 11–14 rows of densely arranged cirri occupy the most postoral area. Keys are provided for all the known species in both genera.     

Two new ciliates from Hong Kong coastal water: Orthodonella sinica n. sp. and Apokeronopsis wrighti n. sp. (Protozoa: Ciliophora)

ABSTRACT. The morphology and infraciliature of Orthodonella sinica n. sp. and Apokeronopsis wrighti n. sp., isolated from the coastal water off Hong Kong, were investigated in living and stained specimens. Orthodonella sinica n. sp. is diagnosed as: a marine Orthodonella 150–310 × 40–80 μm in vivo; with a dominant beak-like projection at the anterior end; about 70 somatic kineties; 84–126 dikinetids in the synhymenium; one contractile vacuole in the posterior one-fourth of cell, near the left margin; one conspicuous dorsal suture. Apokeronopsis wrighti n. sp. is diagnosed as: an Apokeronopsis about 150–230 × 35–55 μm in vivo; dark-reddish blood-cell-shaped cortical granules grouped in three rows on the ventral side and two rows on the dorsal side; 23–35 cirri in the right mid-ventral rows (MVR) and 23–32 cirri in the left MVR; six to eight buccal, two frontoterminal, 30–42 left marginal, and 32–43 right marginal cirri; 21–30 transverse cirri extending anteriorly beyond the level of mid-body; consistently three dorsal kineties. The separation of A. wrighti n. sp. and its highly similar congeners Apokeronopsis crassa and Apokeronopsis bergeri was supported by comparison of their SSrRNA gene sequences.     

Lynnia grapsolytica n. gen,n. sp. (Ciliophora: Apostomatida: Colliniidae), a Deadly Blood Parasite of Crabs with a Novel Pseudocytopharynx

We describe a new genus and species of blood-dwelling apostome ciliate, Lynnia grapsolytica n. gen., n. sp. (Apostomatida: Colliniidae). A distinct kinety “hook” pattern on the tomite’s posterior ventral face, coupled with its marine habitat and use of a decapod host, readily distinguishes this ciliate from all known colliniids. We detected the parasite in ~12% of Pachygrapsus crassipes (Brachyura: Grapsidae) crabs in a California estuary and confirmed its presence at a Baja California rocky intertidal site. As existing methods failed to adequately stain this ciliate, we developed a new miniaturized silver carbonate impregnation staining method that produced excellent somatic and nuclear stains in all five observed cell types. A possibly unique trait is the active invagination of the tropho-tomont’s anterior to form a temporary “pseudocytopharynx,” likely used for feeding. Histological examination revealed that the ciliate invaded and damaged skeletal muscle, the heart, connective tissues, and gonads. Survivorship analysis indicated that infected crabs experienced 2.6 times greater daily mortality than uninfected crabs. Laboratory and field experimental infection attempts failed, suggesting a complex life cycle with outside-host development. Phylogenetic analysis at the 18S and COI loci confirmed the ciliate's placement in the Colliniidae. We emend the diagnosis of Family Colliniidae.     

Urotricha psenneri n. sp. and Amphileptus piger (Vuxanovici, 1962) n. comb., two planktonic ciliates (Protozoa, Ciliophora) from an oligotrophic lake in Austria

Two euplanktonic ciliates, Urotricha psenneri n. sp. (Prostomatida) and Amphileptus piger (Vuxanovici, 1962) n. comb. (Pleurostomatida), were discovered in the surface plankton of the oligotrophic Lake Traunsee in Austria. Their morphology and infraciliature were studied in live cells as well as in specimens impregnated with protargol and silver nitrate. Urotricha psenneri is a small urotrichid, less than 50 microm length and with a single caudal cilium. It is unique in having (i) a massive oral basket projecting as a conspicuous bulge with cylindrical microfibrillar annulus and (ii) a curved brosse row 1 in the broad, barren circumoral area. Amphileptus piger (Vuxanovici, 1962) is about 55 x 13 microm in vivo, has two macronuclear nodules with a single micronucleus in between in the posterior body half, has a single contractile vacuole with a terminal excretory pore, and few, but thick and thus highly conspicuous extrusomes. The amphileptid ciliary pattern (spica) is difficult to recognise due to the widely spaced basal bodies.     

Discovery of a ciliate parasitoid of euphausiids off Oregon, USA: Collinia oregonensis n. sp. (Apostomatida: Colliniidae)

An apostome ciliate, Collinia oregonensis n. sp., is reported inhabiting the cephalothorax and abdomen of 3 euphausiid species from the Oregon-Washington coast: Euphausia pacifica Hansen, 1911, Thysanoessa spinifera Holmes, 1900, and Thysanoessa gregaria G.O. Sars, 1883. This ciliate is the 7th species described for the genus Collinia and the 2nd species known to infect euphausiids. Disease progression and ciliate morphology are described using (1) modified protargol stain, (2) hematoxylin counterstained with Fast Green, and (3) Scanning Electron Microscopy (SEM). All endoparasitic developmental stages (trophont, tomont, tomitogenesis, protomite, and tomite) of C. oregonensis are astomatous and possess between 14 and 22 kineties. C. oregonensis is smaller than C. beringensis Capriulo & Small, 1986, which infects the euphausiid Thysanoessa inermis Kr?yer, 1846 in the Bering Sea and which possesses between 24 and 80 kineties. The ciliate is a parasitoid because it must kill the host to complete its life cycle. Infections and mortalities in multiple host species likely reflect the virulent nature of the ciliate. Adult euphausiids infected with this parasitoid possess a swollen and bright orange cephalothorax. C. oregonensis feeds and proliferates inside euphausiids, producing fulminating infections that rupture the cephalothorax and release large numbers of tomites into the surrounding water. After several hours in the free swimming stage under shipboard conditions in the present study, the tomites adhered to each other, forming filaments. Infection rates ranged between 3 and 20% within individual euphausiid aggregations, but infected aggregations were randomly and sparingly distributed. Infected euphausiids were found at 6.7% of 316 stations sampled during 3 summer cruises. No infected euphausiids were collected in winter. Because E. pacifica and T. spinifera account for about 90% of the euphausiid standing stock in the northern California Current System, this parasitoid ciliate may have a significant impact on euphausiid population abundance, distribution and secondary productivity.     

Hyalophysa Iwoffi sp.n. from the Fresh-water Shrimp Palaemonetes paludosus and Revision of the Genus Hyalophysa

SYNOPSIS. Hyalophysa Iwoffi sp. n. is described from the shrimp, Palaemonetes paludosus . Its life cycle is like that of other exuviotrophic apostomes. Hyalophysa Iwoffi differs from the type species, H. chattoni , in the details of its infraciliature and the shape of its macronucleus during the trophont stage. Emended diagnoses of the genus, Hyalophysa , and the species, H. chattoni , also are given.     

Taxonomic characterization of two marine peritrichous ciliates, Epicarchesium corlissi n. sp. and Pseudovorticella jiangi n. sp. (Ciliophora: Peritrichia), from northern China

Two new marine peritrich ciliates, Epicarchesium corlissi n. sp. and Pseudovorticella jiangi n. sp., were discovered in mariculture waters on the coast of northern China near Qingdao. Their morphology, infraciliature and silverline system were investigated based on both living and silver-impregnated specimens. E. corlissi is characterized as follows: marine Epicarchesium with dichotomously branched stalk; zooids elongate, approximately 60–70×25–35 μm in vivo; peristomial collar double-folded; macronucleus J-shaped; single, small contractile vacuole ventrally positioned; more than 60 striations between peristome and aboral trochal band, 13–18 from aboral trochal band to scopula; abstomal end of row 1 of infundibular polykinety 3 terminating at same level as rows 2 and 3 of infundibular polykinety 3; rows 2 and 3 of infundibular polykinety 3 much longer than row 1 and converging adstomally with infundibular polykinety 1. The new species P. jiangi is diagnosed as follows: marine Pseudovorticella; zooid inverted bell-shaped, approximately 80×60 μm in vivo and with a broad, flat, thin peristomial collar that measures approximately 90 μm across; pellicle with transparent cortical vesicles; macronucleus J-shaped; number of silverlines between peristome and aboral trochal band 20–24, from aboral trochal band to scopula 9–11; abstomal end of row 1 of infundibular polykinety 3 diverges from the other two rows of this polykinety and ends alongside row 3 of infundibular polykinety 2.     

Morphology,morphogenesis and phylogeny of Anteglaucoma orientalis n. sp. (Ciliophora,Oligohymenophorea), a freshwater hymenostomatid from northeastern China

A new hymenostomatid ciliate, Anteglaucoma orientalis n. sp., isolated from a freshwater pond in Harbin, northeastern China, was investigated using live observation and silver staining methods. Anteglaucoma orientalis is characterized as follows: size in vivo about 50–60 × 30–35 μm; oval body shape; buccal area occupies about 25% of body length; 28–36 somatic kineties; membranelle 1 having six or seven basal body rows, membranelle 2 five to seven rows, and membranelle 3 three rows; single macronucleus with one micronucleus attached. Morphogenesis of the genus Anteglaucoma is revealed for the first time. The main events during binary fission are as follows: morphogenesis begins with proliferation of kinetosomes in the middle part of postoral kinety 1, and kinetosomes of this primordial field multiply and organize to finally form the paroral membrane and membranelles 1–3 of the opisthe; the parental apparatus in the proter does not take part in the stomatogenetic process. Phylogenetic analyses based on SSU rRNA gene sequences show that Anteglaucoma orientalis n. sp. clusters with the type species, A. harbinensis Pan et al., 2017, with full support.     

Morphology,ontogeny and molecular phylogeny of a new urostylid ciliate,Bakuella (Pseudobakuella) guangdongica n. sp. (Ciliophora,Hypotrichia) from southern China

The morphology, morphogenesis, and molecular phylogeny of Bakuella (Pseudobakuella) guangdongica n. sp., isolated from southern China, were investigated. The new species is characterized by a body length of 150–225 μm in vivo; 35–42 adoral membranelles; 3–5 buccal, two frontoterminal, 7–12 transverse and two pretransverse ventral cirri; midventral complex comprised of 10–20 pairs and two rows extending to transverse cirri; posterior part of marginal rows slightly overlapping; colorless cortical granules about 1 μm across, arranged in small groups; soil habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures; (2) in the opisthe, the oral primordium originates apokinetally, some old midventral cirri join the formation of frontoventral-transverse cirral anlagen; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Phylogenetic analyses based on the SSU rDNA sequence suggest the non-monophyly of the genus Bakuella.     

Morphology and molecular phylogeny of two new soil ciliates,Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec. (Ciliophora,Hypotrichia) from Tibet

Two novel hypotrichous ciliates, Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec., isolated from soil in the Lhalu Wetland and Motuo Virgin Forest in Tibet, respectively, were investigated using live observation and protargol staining. Hemiurosomoida warreni nov. spec. strongly resembles the type species H. longa but can be distinguished by its body size in vivo (110–145 × 30–40 μm vs. 50–100 × 18–40 μm), number of adoral membranelles (25–38 vs. 15–22), and numbers of right (29–39 vs. 14–23) and left (26–35 vs. 13–23) marginal cirri, transverse cirri (3 vs. 4 or 5) and macronuclear nodules (4–8 vs. 2). Hemiurosoma clampi nov. spec. is characterized by its vermiform body shape, colourless cortical granules distributed in irregular rows, two macronuclear nodules, three frontal cirri, one buccal cirrus, four frontoventral cirri ranged in a line, two transverse cirri, lacking postoral ventral and pretransverse ventral cirri, and marginal rows that are not posteriorly confluent. Phylogenetic analyses based on SSU rDNA gene sequences suggest that Hemiurosomoida is not monophyletic. A close relationship is revealed between Hemiurosomoida warreni nov. spec., Parakahilella macrostoma, Hemiurosoma clampi nov. spec., and the type species Hemiurosoma terricola. As expected, all these species are classified within the “non-oxytrichid Dorsomarginalia”.