ABSTRACT
The research on ciliates, flagelates and opalinates have been widespread by the utilization of techniques employing silver impregnation (protargol), modified by several authors. However, these are time consuming and its results are variable. The present work is a variant of the technique described by Tuffrau (1964, 1967) showing some adaptations made in our laboratory. The organisms can be preserved by different fixatives (alcoholic Bouin, Stieve's fluid, 2.5% glutaraldehyde and others) and then rinsed in destilled water followed by a fast clarification by 3% sodium hypochloride. If the organism is very sensitive to hypochloride, 4% sodium lauryl sulfate may be used and then washed 3 times in distilled water. The protista can be adhered to the glass slides with Mayer's glycerinated-albumin (1 glycerin vol. to 1 or 2 albumin vol.), diluted in water at a proportion of 1:10 Cv/v., or with 1% polylysine followed by fast washes with distilled water. After the slide preparation, they were covered with a layer of 0,8% Silver proteinate. Right after that, the slide has to be placed in a glass tray lined with moist tissue and covered to prevent the proteinate to dry. The tray was placed in a incubator at 40 degrees - 50 degrees C for 30 minutes. The slides are rinsed for 1 minute. with warm (35 degrees C) distilled water. The development of the material should be done with 0.4% hydroquinone with a maximum incubation time of 1 minute. It should be developed gradually, controlling the silver impregnation intensity by observation under optical microscope. Next, rinse in distilled water for 1 minute, and then, fix in 2,5% Sodium thiosulfate. Rinse the slide for two minutes before dehydrating it in an alcoholic serial 50-100 degrees. Finally rinse the slides in xylene. Mount the slides with Entellan MerckTM or Canada balsam.
Subject(s)
Eukaryota/cytology , Silver Proteins , Silver Staining/methods , Animals , Ciliophora/cytology , Ciliophora/ultrastructure , Eukaryota/ultrastructureABSTRACT
This work points out several specific ultrastructural characteristics of the ciliate Phacodinium metchnicoffi. The somatic infraciliature consists of several segments of aciliferous or partially ciliferous monokinetids or alignments of kinetosomes with a particular structuration. All alignments of kinetosomes in double rows are associated with striated kinetodesmal fibers and to a ribbon of postciliary microtubules. The transverse fibers are observed only from monokinetids showing also kinetosomes associated with a 1-2 ribbon of transverse microtubules. However, only the anterior kinetosome of the monokinetid presents a ribbon of transverse fibers and an isolated microtubule next to triplet 5. The somatic kinetosomes located in the zones surrounding the buccal area are clearly associated with a noded microfibrillar network and with nemadesmata on the remaining part of the cortex. The kinetosomes of adorai organelles show 1-2 transverse microtubules. Each organelle is provided with 4 rows of kinetosomes and on each organelle, the kinetosomes of the anterior row bear postciliary microtubules directed towards the front part of the membranelle. The kinetosomes of the left row are associated with 1 or 2 transverse microtubules (T2) as are the kinetosomes of the other rows. The paroral organelle consists of an internal row of kinetosomes accompanied by a succession of parallel small rows each consisting most often of 3 kinetosomes. Only the kinetosomes of the internal row bear postciliary microtubules and prepostciliary microtubules and also show a noded microfibrillar network connecting their proximal zones. Lastly, the taxonomic relationships between Transitella and Phacodinium are discussed.
ABSTRACT
We describe a new genus of Thigmophryidae which is characterized by the cytoskeleton of the thigmotactic area, the median position of the buccal cavity with paroral and adorai organelles. On the locomotive cortex, the kinetosomes are in pairs and ciliated with a median parasomal sac. With the posterior kinetosome are associated: a) a kinetodesmal fibre, b) a ribbon of postciliary fibres, c) a ribbon of transverse microtubules, d) a transverse tractus, e) a microfibrillar tractus. Two or three microtubules run along each kinety. On the thigmotactic cortex, each posterior kinetosome is associated with a kinetodesmal fibre, a postciliary ribbon, a ribbon of transverse fibres and a transverse tractus. Moreover, a skeletal fibre with periodic structure, at the proximal end of the kinetosomes, runs along the left of each kinety. The endoplasm is full of host cells or organisms of the palleal cavity. The ultrastructure of the ciliate is typical of Scuticociliates, just as the buccal organization, with presence of a scuticus. The paroral membrane is a row of dyads with a development of a microfibrillar network as at the level of the single adoral organelle the kinetosomes of which have postciliary fibres.
