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Morphological and Cytological Observations on Iwo Opalinid Endocommensals of Acanthixalus Spinosus (Amphibia, Anura)

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Morphological and Cytological Observations on Iwo Opalinid Endocommensals of Acanthixalus Spinosus (Amphibia, Anura) See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/238037392 Morphological and cytological observations on two opalinid endocommensals of Acanthixalus spinosus (Amphibia, Anura) Article in Canadian Journal of Zoology · February 2011 DOI: 10.1139/z96-171 CITATIONS READS 2 133 3 authors, including: Félix-Marie Affa'a RAPPAUC 21 PUBLICATIONS 98 CITATIONS SEE PROFILE All content following this page was uploaded by Félix-Marie Affa'a on 23 January 2015. The user has requested enhancement of the downloaded file. 1573 Morphological and cytological observations on Iwo opalinid endocommensals of Acanthixalus spinosus (Amphibia, Anura) Félix-Marie Affa'a, Jean-Pierre Mignot, and Jean-Louis Amiet Abstract: The morphology and cytology of two new opalinid species were studied using silver impregnation and fixation, which preserves the microfibrils. Both species, commensal on Acanthixalus spinosus, are hast-specifie. Light microscopy showed the existence of a posterior secant system in Opalina proteus n.sp. and its absence in Cepedea couillardi n.sp. (in agreement with the differences presently recognised between the two genera). At the ultrastructural level, however, bath species present a posterior fibrillar zone that seems to be homologous with the secant system. This apparent contradiction may be explained by the fact that the secant system is visible under light microscopy only in O. proteus because its fibrillar zone is more developed than in C. couillardi. The life cycle of C. couillardi spans stages from the tadpole to the adult; in contrast, O. proteus completes its cycle before metamorphosis of the hast. Résumé: Les auteurs ont étudié la morphologie et l'ultrastructure de deux nouvelles opalines par imprégnation à l'argent en microscopie optique et en microscopie électronique, après fixation par une technique réputée préserver les microfibrilles. Les deux espèces, commensales d'Acanthixalus spinosus, sont spécifiques de leur hôte. La microscopie optique a montré l'existence d'un système sécant postérieur chez Opalina proteus n.sp. et son absence chez Cepedea couillardi n.sp. (ce qui est conforme à la différence jusqu'ici reconnue entre les deux genres). Cependant, en microscopie électronique les deux espèces présentent une zone fibrillaire postérieure qui paraît correspondre à un système sécant. Cette contraction apparente peut être due au fait que le système sécant est visible en microscopie optique seulement chez Opalina parce que cette zone fibrillaire y est plus développée que chez Cepedea. Au cours de son cycle vital, Cepedea couillardi passe des têtards aux adultes alors qu'O. proteus effectue tout son cycle avant la métamorphose de la grenouille. 1 Introduction here. The general characteristics of these two genera, bath multinucleate, are presented in Table 1. Further details cali Acanthixalus spinosus (Buchholz et Peters 1875) is the sole be round in Affa'a (1992) and Affa'a and Lynn (1994). species in a genus whose morphology and anatomy set it apart frOIDother African tree frogs (Perret 1962). It repro- duces exclusively in tree cavities of sufficent size to hold Materials and methods water year-round (Perret 1962). A few other African anuran Frogs were captured in some 20 localities in the South Cameroon species exhibit such reproductive behaviour: Hyperolius Plateau, south of Yaoundé (11-12°E, 3 - 3.5° N), at altitudes from if acutirostris, H. mosaicus, Nectrophryne afra, and N. batesi 650 to 750 m. During the 5 years (1989-1994) when we specifi- (Amiet 1989). These breed in smalIer cavities that lose their cally looked for opalinids from Acanthixalus spinosus, more than a water in the dry season. 100 tree cavities were prospected and frogs were found in only 28. The unique systematic position and ecology of A. spinosus Ten ofthese 28 were visited many times a year to collect pneumonts calI for particular attention to be raid to ifs endocommensal and tadpoles at different stagesof developmentand at metamorphosis. Protozoa. Thus, one of us was able to describe several species ln total, 136 tadpoles and 59 pneumonts were collected, 1 of the of nyctotherans that are endemic to it (Affa'a 1979, 1980, new species being found in 74 (54%) tadpoles and the other species in 18 pneumonts (30.50%) and 14 tadpoles (10.29%). 1986, 1988). The abject of the present parer is to describe ifs The difficulty in finding hasts, the scarcity of one of their endozoic opalinids. Only two species, belonging to the genera opalinids, and the difficulty in redreating in the laboratory the partic- Cepedea and Opalina, were round, and they are described ular conditions oftree cavities colonized by A. spinosus or offinding a medium suitable for cultivating opalinids did not allow us to Received August 8, 1995. Accepted February 23, 1996. follow the developmental life cycle of the species described here. F.-M. Affa'a.1 Département de biologie, Université The presence of adults and tadpoles of A. spinosus in the tree cavi- d'Ottawa, 30 Marie Curie, Ottawa, ON KIN 6N5, Canada. ties year-round, however, allows us to propose that transmission J.-P. Mignot. Laboratoire de biologie des protistes, Unité de of Protozoa to very young tadpoles occurs when cysts contained in recherche associée n° 1944, Université Blaise-Pascal de the faeces of pneumonts and aider tadpoles are swallowed. Clermont-Ferrand II, 63177 Aubière Cédex, France. Preparation of opalinids by silver impregnation for light micros- J.-L. Amiet. 48, rue des Souchères, 26110, France. cary (Affa'a 1979, 1986, 1988, 1991, 1992) and special fixation procedures for ultrastructural studies of the fibrillar system (Mignot Author to whom ail correspondence should be addressed 1994; Mignot and Affa'a 1994a, 1994b, 1994c) has been previously (e-mail: [email protected]). described. Cano J. Zoo\. 74: 1573-1584 (1996). Printed in Canada / Imprimé au Canada 1574 Cano J. Zool. V 2 3 4 1 Affa'a et al. 1575 Figs. 1-5. Cepedeacouillardi n.sp., ammoniacalsilver nitrate impregnationsafter Fernandez-Galiano(1976).Fig. 1. Mature individual as found in pneumonts. Figs. 2 and 3. Sigmoid or arcuate immatures from young tadpoles. Fig. 4. Anterior part of a mature individual. Scale bars = 50 Jtm. Fig. 5. Anterior zone of a mature individual seen from the right lateral side, showing the semicircular falcu1ar zone. Scale bar = 20 Jtm. Table 1. Morphological differences between Opalina and Cepedea. Opalina Cepedea Body shape Wide, leaflike Elongated, narrow, round, or flattened Caudal secant system and (or) kinety Present Absent coalescence zone at posterior pole Ratio of length of falcular zone to >40% <25% cell perimeter Results Fig. 6. Outline drawings of the various forms of Cepedea couillardi from Protargol-impregnated individuals. Endozoic organisms and host specificity The commensal fauna of the lower intestine of Acanthixalus spinosus varies with the age of the hast and, to a lesser extent, with microhabitat and locality. Opalinids have been found associated with diverse Metazoa such as nematodes (espe- cia11yoxyurids), which occur in adults as we11as in tadpoles. Rotifers were found only in tadpoles froID4 of the tree cavities sampled. Protozoa are far more abundant, as bath species and individuals. Six species of nyctotherids have been described (Affa'a 1979, 1980, 1986, 1988), and a further species has yet to be described. One unspecified Trichodina species has also been found in tadpoles froID two sites. Nyctotheroides augeri is transmitted frOIDtadpoles to adults in the course of its life cycle, the others being restricted to the larval stage of the hast. Proteromonads, diplomonads, and (or) oxymonads and trichomonads are found in bath tadpoles and adults. ln the latter, they coexist with one undetermined Balantidium species and three nyctotherids. Both opalinid species described here differ from a11those observed during a study of the endocommensal Protozoa of 100 anuran hast species froID different location, primarily Cameroon. They also differ froIDthose described by Metcalf (1923, 1940) froIDthe Afrotropical region. ln particular, we have searched for these new opalinid species in Hyperolius acutirostris, a frog that breeds in the saille tree cavities as A. spinosus, known to be a hast ofthese opalinids. Examina- tion of20 tadpolesand 15adults of H. acutirostrisrevealedthe presence of one opalinid, Cepedea affinis, and two nyctho- terids, one specimen of Albaretia terroni and two of Albaretia maxima. It thus seems that our two new opalinids are oioxenous, that is, they are restricted to a single hast (Euzet and Combes 1980), as are the numerous nyctotherid species 1 ribbonlike (Figs. 1 and 6) with a wide and latera11yflattened commensal on A. spinosus (Affa'a and Amiet 1990, 1994). anterior extremity (Figs. 4 and 5) and a minute caudal tir. The numerous nuclei are difficult to stain by conventional Cepedea couillardi n.sp. techniques. The falcular zone is subapical and slightly dis- Figs. 1-15 placed to the right side. It often takes the form of a half-circle Morphological observations with moderatelytapered extremities. Kinetiesoriginate directly Mature trophonts of this opalinid, which are very large frOIDthis zone. They mn straight on the right side and curve (Table 2), have so far been found only in adult frogs and around to lie more or Jess diagona11yon the left surface. young pneumonts (Affa'a et Amiet 1994) in 30% (18/59) of Immature forfis, found exclusively in tadpoles (14/136, the A. spinosus population sampled. They are elongated and or 12.35%), are sma11erand variable in shape. The sma11est, 1576 Cano J. Zool. Vol. 74, 1991 Affa'a et al. 1577 Figs. 7 and 8. Cortex of Cepedea couillardi. Tangential sections of the cortex at various levels, showing the superficial cytoskeletal network. Fig. 7. 21000 x. Scale bar = 1 pm. Fig. 8.42000 x. Scale bar = 0.5 /lm. Edv, endocytotic vesicles; Exv, exocytotic vesicles; F, fibrillar bundles. Ks, kinetosomes of somatic flagella. Fig. 9. Ectoplasm and endoplasm of Cepedea couillardi at low magnification.
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