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Three species of the genus Trichodina Ehrenberg, 1838 and one species of the genusChilodonella Strand, 1926 were collected from gills of Carassius auratus. They areTrichodina luzhoues sp. n., Trichodina mutabilis Kazubski and Migala, 1968, Trichodina reticulata Hirschman and Partsch, 1955, and Chilodonella hexasticha Kiernik, 1909. T. luzhoues sp. n. is a medium-sized trichodinid, and its denticles are very distinctive: the blade is narrow rhombus shaped, the section connecting the blade and central part is long and very thin; the section connecting the central part and ray is short and very thick. Ch. hexasticha is a new record in China.
Parasitol Res (2012) 111:433–439 DOI 10.1007/s00436-012-2859-0
Ciliate ectoparasites (Ciliophora: Trichodinidae/ Chilodonellidae) on gills of Carassius auratus from the Yangtze River, China, with the description of Trichodina luzhoues sp. n.
Received: 14 July 2011 / Accepted: 7 February 2012 / Published online: 29 February 2012 # Springer-Verlag 2012
Abstract Three species of the genus Trichodina Ehrenberg, 1838 and one species of the genus Chilodonella Strand, 1926 were collected from gills of Carassius auratus. They are Trichodina luzhoues sp. n., Trichodina mutabilis Kazubski and Migala, 1968, Trichodina reticulata Hirschman and Partsch, 1955, and Chilodonella hexasticha Kiernik, 1909. T. luzhoues sp. n. is a medium-sized trichodinid, and its denticles are very distinctive: the blade is narrow rhombus shaped, the section connecting the blade and central part is long and very thin; the section connecting the central part and ray is short and very thick. Ch. hexasticha is a new record in China.
About ten trichodinds have been found from Carassius auratus so far, such as Trichodina oviformis Chen, 1955, Trichodina nobilis Chen, 1963, Trichodina carassii Li, 1990, Trichodina paranigra Tong, Zhao and Chen, 2004, Trichodina pachyhamata Tang and Zhao, 2005, Trichodina subtilihamata Tang, Zhao and Tao, 2007, Trichodina bev- icirra Tong and Zhao, 2010, and so on. Two Chilodonellids, Chilodonella cyprini (Hofer 1906; André 1912; Bespalyj 1950; Krascheninnikow 1952; 1953; Chen 1955; Smirnova et al. 1964; Grabda 1971) and C. hexasticha (André 1912), have been reported from C. auratus.
The present paper deals with three ciliates belonging to the genus Trichodina Ehrenberg, 1838 and one ciliate of the genus Chilodonella Strand, 1926 collected from gills of C.
Y. Hu (*) Luzhou Vocational & Technical College, Luzhou 646005 Sichuan, People’s Republic of China e-mail: firstname.lastname@example.org
auratus. They are Trichodina luzhoues sp. n., Trichodina mutabilis Kazubski and Migala, 1968, Trichodina reticulata Hirschman and Partsch, 1955 and Chilodonella hexasticha Kiernik, 1909.
Materials and methods
The host C. auratus (L.) that was more than 1 year old was obtained in 2009 from the Yangtze River at Luzhou, Sichuan, China (28°51′N, 105°23′E), which is the longest river in China.
Gill scrapings were made from the hosts. Smears with ciliates were air dried and then the slides with ciliates were impregnated with Klein's dry silver impregnation technique (Klein 1958). The nuclear apparatus was shown using methyl green-pyronin staining (Foissner 1991). All photomicro- graphs and illustration drawings were made with the help of a camera (Motic DM–BA300–B) at ×100 magnification with oil immersion lens and software Motic Images Advanced 3.2 and CorelDraw X3. The statistics were obtained with Micro- soft Excel 2003.
All measurements are presented in micrometers (μm). In each case, minimum and maximum values are given, followed in parentheses by the arithmetic mean and standard deviation. In the case of the number of radial pins/denticle and denticles of the trichodinid and number of kineties of the chilodonellid, the mode is given rather than the arithme- tic mean with the number of specimens examined given in parentheses. The body diameter of trichodinid is measured as the adhesive disc plus the border membrane. The meas- urements of trichodinid follow the uniform specific charac- teristics proposed by Lom (1958) while the method proposed by Van As and Basson (1989) was followed for denticle description, as shown in Fig. 1.
Parasitol Res (2012) 111:433–439
Fig. 1 Diagram to illustrate denticle structure and construction of X- and Y-axes as fixed references for description of denticles (after Van As and Basson 1989). AB, apex of blade; AM, anterior margin of blade; AR, apophysis of ray (0 thorn); B, blade; BA, apophysis of blade; CA, central of adhesive disc; CB, section connecting blade and central part; CC, section connecting central part and ray; CCP, central conical part; CP, central part; DC, deepest point of curve; DS, distal surface of blade; PM, posterior margin of blade; PP, posterior projection; PR, point of ray; R, ray; SA, section of central part above X-axis; SB, section of central part below X-axis; TP, tangent point
The position of the micronucleus of trichodinid is given relative to the arch-shaped macronucleus, according to the format described by Lom (1958), which was based on the system originally proposed by Dogiel (1940). In this system, the micronucleus is situated in one of three positions relative to the terminations of the arms of the macronucleus: (1) externally, near the right termination (+y); (2) externally, between the two terminations (−y); and (3) internally, near the right termination (−y1).
Trichodina luzhoues sp. n.
Species: T. luzhoues sp. n. Family: Trichodinidae Claus, 1874 Type Host: Carassius auratus (L.) Fish Family: Cyprinidae Type Locality: Luzhou, Sichuan, China (28°51′N, 105°23′E) Location: Gills of Carassius auratus (L.) Date of Sampling: 7/2009 Etymology: The specific epithet “luzhoues” is coined
from the name of “Luzhou,Sichuan, China”. Reference Material: Holotype, slide LZY109/2009, and
paratype slides LZY112/2009, LZY113/2009 are deposited in the Biological Laboratory of Luzhou Vocational & Tech- nical College.
The following is a description of a medium-sized, freshwater trichodinid:bodydiameter,36.6–61.0(50.3 ± 7.6);adhesive disc,28.1–48.7(40.5 ± 6.6)(seeFigs.2–5,6;Table1);dentic- ulatering,18.1–31.2(25.2 ± 4.7);bordermembrane,4.1–6.2 (4.9 ± 0.6);denticlenumber,21–29(25);radialpinsper
Figs. 2–5 Photomicrographs of T. luzhoues sp. n. from C. auratus, after dry silver impregnation (2, 3, 4) and green-pyronin staining (5). 2–3, adhesive discs; 4, adoral ciliary spiral; 5, nuclear, MA macronucleus; MI micronucleus (scale bars020 μm)
Results and discussions
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denticle,5–6(5);denticlespan,9.7–17.1(13.3 ± 2.6);denticle length,4.1–7.7(6.0 ± 1.2);thecentralzoneofadhesivedisc clear; the blade is narrow rhombus shaped; and blade length, 3.0–7.8(5.3 ± 1.3).Distalsurfaceofbladeisshort,tangentpoint rounded. The anterior margin of blade is approximately parallel to curves of posterior margin of blade, forming almost L shape; apexofbladetouchesY + 1-axis,somecrossesY + 1-axis,apoph- ysis of blade barely seen; section connecting the blade and central part is long and very thin. The central part is well developed with rounded point fitting tightly into preceding denticle,extendingabouthalftowardstheY − 1-axis.Sectionof central part above X-axis is less than the section of central part below X-axis in shape. The central part width is 1.2–4.5 (2.3± 0.7). The section connecting central part and ray is very thick. The ray is directed towards the center of adhesive disc. The point of the ray is sharp or rounded. Ray length is 3.7–8.3 (5.7±1.3). Adoral ciliary spiral makes a turn of 407–420°. Macronucleus is horseshoe shaped, elongated, and with characteristic dilations at both ends; external diameter is 31.3–35.1 (33.2±1.8). Micronu- cleus is spherical; diameter is c.3.3, situated in −y1 position.
About ten trichodinds have been found from C. auratus so far, but T. luzhoues sp. n. is obviously different from them (see Figs. 6–8 and Table 1). T. luzhoues sp. n. only resembles Trichodina cooperi Poynton and Lom, 1989 obtained from skin, fins of Gadus morhua L. found in Nova, Scotia, Cana- dia, and Trichodina galyae Lom and Laird, 1969 obtained from the gills of Cyelopterus lumpus found in Canada.
T. luzhoues sp. n. is different from T. galyae by the shape of denticle and some other measurements. (1) The new species is smaller than T. galyae, for example, body diam- eter (36.6–61.0 vs. 70–85), adhesive disc (28.1–48.7 vs. 48–65), and denticulate ring (18.1–31.2 vs. 30–42). (2) In the case of the new species, the posterior tip of the central part extends almost halfway to Y−1-axis, but in the case of T. galyae, it almost touches Y−1-axis. (3) The ray is thicker in the new species than in T. galyae. (4) Morphometic data of the new species also varies when they are compared with those of T. galyae, e.g., radial pins per denticle (5–6 vs. 10–12), denticle length (4.1–7.7 vs. 10.5; see Table 1).
Figs. 6–8 Diagrammatic drawings of the denticles of trichodinid cil- iophorans.6, T. luzhoues sp. n.; 7, T. galyae; 8, T. cooperi
T. luzhoues sp. n. is clearly distinguished from T. cooperi too. (1) The new species is smaller than T. cooperi, i.e., body diameter (36.6–61.0 vs. 95–122), adhesive disc (28.1–48.7 vs. 82–107), and denticulate ring (18.1–31.2 vs. 49–67). (2) The shapes of blade are different in the two trichodinids. The section connecting the blade and the central part is very thin in the new species but thick in T. cooperi. (3) The ray parallels Y-axis and directs towards the center of adhesive disc in the new species, but it slants backward and is slightly forward curved in T. cooperi. (4) Morphometic data of the new species also varies when they are compared with those of T. cooperi, for instance, border membrane (4.1–6.2 vs. 6.4–8.9), radial pins per denticle (5–6 vs. 7–9) denticle span (9.7–17.1 vs. 23.6–32.5), denticle length (4.1–7.7 vs. 20.4–24.5), blade length (3.0–7.8 vs. 7.5–10.0), central part width (1.2–4.5 vs. 3.2–6.3), ray length (3.7–8.3 vs. 10.2–19.1), and adoral ciliary spiral (407–420 vs. 370–390; see Table 1).
T. mutabilis Kazubski and Migala, 1968
The following is a description of a large-sized freshwa- ter trichodinid (see Figs. 9–10 and Fig. 13): body di- ameter, 77.5–97.0 (84.4±5.8); adhesive disc, 61.3–77.0 (67±4.3); central area is not clear; denticulate ring, 38.1–49.1(42.3 ± 3.0);bordermembrane,5.8–9.9 (8.3±1.1); denticle number, 25–27 (27); and radial pins per denticle, 6–7 (6). Blade is prismatic, long, and narrow. Tan- gent point is sharp. Distal surface of blade is straight and lower than tangent point. Anterior margin of blade is forming arch curve, parallel to posterior margin of blade, apex of blade touching Y+1-axis, and apophysis of blade is barely seen. Central part is narrow with rounded point fitting tightly into preceding denticle, almost extending half way to Y−1-axis. Shapes of the central part above and below X-axis are dissim- ilar. Width of central part is 1.0–1.5 (1.3±0.2). Ray is slim, straight, and slanted forward, forming an angle of about 30° with Y+1-axis. Adoral ciliary is spiral, 410–430°.
T. mutabilis was originally described by Kazubski and Migala from Poland in 1968. Since then, T. mutabilis has been reported from various places in Eastern Europe, For- mer USSR, South Africa, Israel, and India (Lom 1970; Migala 1971; Kashkovsky 1974; Jusupov and Urazbaev 1980; Basson and Van As 1994; Mitra and Bandyopadhyay 2005; Dove and Donoghue 2005).The denticles of my ma- terial resemble those reported by Kazubski and Migala (1968) in winter.
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Figs. 9–12 Photomicrographs of the silver nitrate impregnated of Trichodina spp. 9–10, T. mutabilis Kazubski and Migala, 1968; 11–12, T. reticulata Hirschman and Partsch, 1955 (scale bars020 μm)
T. reticulata Hirschman and Partsch, 1955
The following is a description of a middle-sized freshwater trichodinid, cap shaped (see Figs. 11–12 and Fig. 14): body
diameter,47.7–52.3(49.9 ± 1.8);adhesivedisc,36.9–40.8 (38.7±1.5); denticulate ring, 24.0–27.6 (25.7±1.1); border membrane,10.8–11.6(11.2 ± 0.3);denticlenumber,22–25 (24); radial pins, per denticle 9–11 (10); denticle length, 10.8–13.5(12.2 ± 0.9);denticlespan,5.1–7.1(6.1 ± 0.6);cen- tral area with granules consists of 13–16 (14) cell-like
Table 1 Morphometric comparison of T. luzhoues sp. n. and T. galyae, T. cooperi (measurements in micrometers)
Host Locality Site References No. of specimens measured Body diameter
Adhesive disc Denticulate ring Border membrane Denticle number Radial pins/denticle Denticle span Denticle length Blade length Central part width Ray length
Figs. 13–14 Diagrammatic drawings of the denticles of trichodinid ciliophorans 13, T. mutabilis; 14, T. reticulata
structures, giving a reticulated appearance. Tangent point is distinctly round and head of blade is almost bulbous at this point. Anterior margin of blade is parallel to curves of posterior margin of blade; apex of blade extends beyond Y +1-axis; posterior margin of blade is with deep curve, form- ing almost C-shape; apophysis of blade is unobvious; sec- tion connecting blade and central part is thin and short. Bladelengthis3.4–6.2(5.1 ± 0.8).Thecentralpartiswell developed, short and stout, of same thickness throughout, extending slightly only beyond Y-axis, and fitting tightly into preceding denticle with blunt rounded point. The
section of central part above and below X-axis is similar in shape, central part width is 1.7–2.5(2.1±0.3). The section connecting central part and ray is very short, the apophysis of ray is sharp, the ray is thick, the point of ray is blunt rounded, and ray length is 4.4–5.5(5±0.4). Adoral ciliary is spiral, 400–410°.
T. reticulata was originally described by Hirschmann 1955. T. reticulata has been reported from Former USSR, Eastern Euope, South Africa, Asia, and the USA (Lom 1960; Chen 1963; Lom and Hoffman 1964; Lom et al. 1976; Grigoryan and Stein 1981; Kazubski 1982, 1988; Basson et al. 1983; Albaladejo and Arthur 1989; Basson and Van As 1993). This species is clearly recognizable based on the distinct shape of the denticles, and granules as cell-like structures in the central area. T. reticulata described in this paper repre- sents one of the lowest ranges of body dimensions reported so far. It was reported by Chen in China in 1963, but photomicrographs of the trichodinid were not provided.
Chilodonella hexasticha Kiernik, 1909
The chilodonellid body is flattened at the ventral (oral) side, the dorsal side is rounded in shape (see Figs. 15–18). Body
Figs. 15–18 Photomicrographs of the silver nitrate impregnated of C. hexasticha (scale bars020 μm)
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length is 60.5–91.6 (75.5±2.4)μm, width is 53.9–77.8 (62.2± 1.7)μm. The ciliature of the ventral body side is composed of a short preoral kinety and two systems of kineties. The left and right system of kineties is conspicuously separated by a gla- brous area. In the anterior part of this zone is the oral opening. Kineties is loosely arranged and the distances between them is not equal. The right system consists of 7–9 (8) kineties and 1–3 (2) postoral kineties. The two outermost rows begin more anteriorly surrounding at front of the kineties of the left system. The left system consists of 7–8(7) kineties. In the system are short rows at the inner side in the posterior as well as outer side in the anterior part of the body. Others exist between the two kineties.
Macronucleus is large and oval (see Fig. 16), length is 10.2–12.8(11.5±1.2)μm, width is 7.5–8.9 (8.2±1.3)μm. Micronucleus is not visible.
Chilodonella cyprini and Ch. hexasticha were described in the first decade of twentieth century. Ch. hexasticha has been reported from Former USSR, Germany, Poland, Czechoslovakia, and the USA (Kiernik 1909; Prost 1952; Kazubski and Migala 1974; Lom et al. 1976; Wierzbicka 1997). However, descriptions of Ch. cyprini and Ch. hexasticha were not precise due to imperfect methods of study used at that time, so the distinctness of both species was questioned, before Kazubski and Migala (1974) described Ch. cyprini and Ch. hexasticha in detail and expounded the differences between them. There are differences between Ch. cyprini and Ch. hex- asticha by Kazubski and Migala: (1) These species differ mainly by the number of kineties, which is larger in Ch. cyprini and smaller in Ch. hexasticha. (2) The differences concern the arrangement of kineties. In Ch. cyprini, the kineties are close one to the other, lying in nearly equal distances, while in Ch. hexasticha the kineties are loosely arranged and the distances between them are not equal. Especially the inner kineties of the right system are outstanding, lying in much greater distances then the other kineties of both systems. (3) There are also some differences in the number of the postoral kineties (5–6 in Ch. cyprini and only 1–3 in Ch. hexasticha). (4) Ch. cyprini most frequently occurs on young fishes, on their gills and skin, while Ch. hexasticha occurs usually on gills of older fishes.
The chilodonellid in this paper is obvious Ch. hexasticha, but its body size is larger than that of individuals in other populations of Ch. hexasticha. It is a new record in China.
AcknowledgementsThis work was supported by the Natural Science Foundation of Luzhou (project no. 06112 and project no. 2010-S-21). I
would like to thank Dr. Chengwen Li for his excellent technical assistance in the laboratory.
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