Silver Carbonate Impregnation Research Articles

First record of Colpodidium caudatum (Ciliophora, Colpodidiidae) in Europe

The present study documents the discovery of the first European population of Colpodidium caudatum (Ciliophora, Colpodidiidae) in a water drain in a school playground in Manzanares (Ciudad Real, Spain). This species has been documented on every continent except Antarctica and Europe, until now. The ciliate was isolated from wet run-off soil collected from the water drain and was grown in semi-permanent cultures in the laboratory. The infraciliature of the ciliate was revealed using silver carbonate impregnation and cell measurements were taken from living and silver-impregnated specimens. A comparative analysis of published data from various populations of C. caudatum across the globe showed high intraspecific morphological variability in this species. To differentiate between species within the Colpodidium genus, a dichotomous key is presented. This investigation shows that C. caudatum is a ciliate that is found all over the world and is particularly associated with terrestrial habitats that are periodically flooded.

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.

Morphological and Morphometric Characterization of Eodinium posterovesiculatum (Ciliophora, Ophryoscolecidae) in Brazilian Cattle

The morphological variability of Eodinium posterovesiculatum (Ciliophora, Trichostomatia) has been interpreted in different ways: four distinct species or four morphotypes of the same species. The present study aims to perform morphological and morphometric comparative analysis of the four morphotypes found in cattle from the southeastern region of Brazil. Ruminal content samples were obtained from four Holstein x Gir cattle and fixed at 18.5% formalin for morphological analysis. Morphometry was performed based on individuals stained with Lugol's solution [1]. The infraciliary bands were stained using silver carbonate impregnation technique [2]. Morphological and morphometric characterizations, supported by literature, suggest that the four morphotypes of E. posterovesiculatum are actually a single polymorphic species due to small morphometric differences and mostly identical morphological characters, with the format of the caudal processes being the only morphological characteristic that sets them apart.

Infraciliature of Eudiplodinium dilobum, E. rostratum and E. maggii

The aim of this study was to demonstrate the infraciliature of rumen ciliates, Eudiplodinium dilobum , E. rostratum , and E. maggii by using pyridinated silver carbonate impregnation method. The specimens of E. dilobum , E. rostratum , and E. maggii were obtained from the rumen contents of water buffaloes ( Bubalus bubalis ) in Kastamonu, Turkey. As the result of the investigation, Diplodinium -type infraciliature was detected in E. dilobum , E. rostratum, and E. maggii . The buccal infraciliature of E. dilobum , E. rostratum , and E. maggii was composed of two polybrachykineties and paralabial kineties. The adoral polybrachykinety encircled most of the circumference of the vestibular opening and the vestibular polybrachykinety extended along the dorsal wall of the vestibulum from the inner edge of the adoral polybrachykinety.

Morphological Features of Bundleia spp. (Ciliophora: Entodiniomorphida: Buetschliidae) and Ontogenesis in Bundleia Cunha and Muniz 1928

The aim of this study was to investigate the morphological features of 7 endocommensal ciliates, B. (s. str.) postciliata, B. (s. str.) piriformis, B. (s. str.) nana, B. (C.) elongata, B. (C.) triangularis, B. (F.) dolichosoma, and B. (F.) inflata belonging to the genus Bundleia, which exist in the hindgut of 15 Rahvan horses and 15 racing horses living in the vicinity of Izmir and to investigate the ontogenesis in the Bundleia genus. Specimens were obtained from 30 horses and were fixed with 10% formalin. The morphological features of the species were investigated with the pyridinated silver carbonate impregnation and silver nitrate impregnation techniques at the level of light microscopy. The morphological features of Bundleia spp. were found to be similar to the original descriptions, and it was detected that ontogenesis in the Bundleia genus exhibited suitability with other ciliate species in the Entodiniomorphida order. We conclude that the geographical variation and feeding habits of host animals are important for evaluating the biometric data of species.

Methods for taxonomic studies of rumen ciliates (alveolata: ciliophora): a brief review.

This review presents the principal methods used in taxonomic studies of rumen ciliates: live observation, Lugol staining, fixation and staining with methyl-green formalin saline (MFS) solution, protargol staining, silver carbonate impregnation, scanning electron microscopy and molecular techniques. Mastering these techniques is essential for successful research on the taxonomy of rumen ciliates. No single technique reveals all of the characteristics required for a complete description of a rumen ciliate; therefore, it is necessary to combine the use of these techniques as appropriate to the rumen ciliate group under study. Tables are provided to summarize: 1) morphological methods more appropriate for revealing morphological structures of interest, 2) morphological methods indicated for each group of rumen ciliates, and 3) main primers used for PCR amplification of the 18S rDNA of rumen ciliates.

An update of ‘basic light and scanning electron microscopic methods for taxonomic studies of ciliated protozoa’

This is an update of the review by Foissner (1991). Since then, I have improved some methods considerably. The following methods are described in detail: live observation, supravital staining with methyl green-pyronin, dry silver nitrate impregnation, wet silver nitrate impregnation, silver carbonate impregnation, protargol impregnation (three procedures), scanning electron microscopy, and deciliation. Familiarity with these methods (or modifications) is a prerequisite for successful taxonomic work. No staining method is equally appropriate to all kinds of ciliates. A table is provided which indicates those procedures which work best for certain groups of ciliates. A second table relates to the structures revealed by the procedures. Good descriptions usually demand at least live observation, silver nitrate and protargol or silver carbonate impregnation. Some instructions are provided for distinguishing mono- and dikinetids as well as ciliated and non-ciliated basal bodies in silvered ciliates. Furthermore, I added a section on 'Deposition and Labeling of Preparations'. All methods work not only with ciliates but also with many other heterotrophic and autotrophic flagellated and amoeboid protists. The brilliancy of silver preparations has tempted some taxonomists to neglect live observation. However, many important species characteristics cannot be seen or are changed in silvered specimens. I thus consider all species descriptions based exclusively on silver slides as incomplete and of doubtful value for both α-taxonomists and ecologists.

Occurence of the Hindgut Ciliates Paraisotricha colpoidea and P. minuta (Ciliophora: Paraisotrichidae) in Horses in Turkey

The aim of this study was to determine the cytological features of 2 endocommensal ciliates, P. colpoidea and P. minuta belonging to genus Paraisotricha found in the hindgut of 15 Turk rahvan and 15 English and Arabic horses from Izmir, compare the morphological characters of species with their original descriptions and previous reports and discuss the similarities and differences. The cytological features of two endocommensal ciliates were investigated with the pyridinated silver carbonate impregnation and silver nitrate impregnation techniques at the level of light microscopy. Specimens from our horses were found to be similar to the original descriptions on the basis of cytological features, morphological characters and biometric data. We have concluded that the geographical variation and feeding habits of the host animals are important for evaluating the biometric data of species.

First record and redefinition of the Qingdao population of marine ciliate Cardiostomatella vermiformis (Kahl, 1928) Corliss, 1960 (Protozoa, Ciliophora)

The living morphology and infraciliature of a marine ciliate, Cardiostomatella vermiformis isolated from a sand beach of the Jiaozhou Bay near Qingdao, China, were investigated by live observation, protargol and silver carbonate impregnation methods. Both the morphological and morphometric data largely agree with former records. Based on the data obtained, an improved diagnosis for Cardiostomatella vermiformis is supplied: Large marine Cardiostomatella with cylindrical body shape and several prolonged caudal cilia; cells in vivo about (90–500)μm × (30–120)μm; macronucleus beaded; single contractile vacuole caudally positioned; buccal apparatus conspicuously small, genus typical; three to six postoral kineties; 96–130 somatic kineties. This species is recorded for the first time in China.

Descriptions of new Parentodinium ciliates in the family Parentodiniidae n. fam. from Hippopotamus amphibiusin comparison with some entodiniomorphs from horses and cattle

Descriptions of Parentodinium ostrea Thurston & Noirot-Timothee, 1973 and P. africanum Thurston & Noirot-Timothee, 1973 are revised on the basis of observation by pyridinated silver carbonate impregnation. Two new species, Parentodinium pediculum n. sp. and P. lituus n. sp., are also described. Based on the presence of caudal lobes, P. ostrea is divided into three forms, f. ostrea Thurston & Noirot-Timothee, 1973, f. euostrea n. f., and f. ablobum n. f. and P. pediculum is divided into two forms, f. pediculum n. f. and f. dorsale n. f. Until now, Parentodinium has been included in the family Cycloposthiidae. Because of its characteristic arrangement of infraciliary bands, a new family Parentodiniidae is proposed. Parentodinium ciliates are compared with five entodiniomorphid ciliates, Bundleia postciliata, Blepharocorys valvata, and Cycloposthium bipalmatum from the intestine of horses and Entodinium simplexand Eudiplodinium maggii from the rumen of cattle. Parentodinium has characteristic features intermediate between other entodiniomorphid families in both general and infraciliary morphology. Parentodinium has an ovoidal body laterally compressed, an ovoidal macronucleus varying in location, a micronucleus adhering to various sides of the macronucleus, and a single contractile vacuole in a posterior ventral location. Parentodium ostrea, P. africanum and P. pediculum have a cytoproctal ciliary tuft. Parentodinium has a characteristic arrangement of infraciliary bands distinct from other entodiniomorphid ciliates. The adoral polybrachykinety completely encircles the circumference of the vestibular opening. There are several vestibular kineties extending along the dorsal wall of the tubular vestibulum. Paralabial kineties are located at the right ventral side of the adoral polybrachykinety. Cytoproctal kineties run inside the cytoproct.

Morphology and Morphogenesis of Strobilidium caudatum (Fromentel), Meseres corlissi N. Sp., Halteria grandinella (Müller), and Strombidium rehwaldi N. Sp., and a Proposed Phylogenetic System for Oligotrich Ciliates (Protozoa, Ciliophora)1

ABSTRACT. The morphology and morphogenesis of some oligotrichs were investigated using protargol impregnation, silver carbonate impregnation and scanning electron microscopy. The somatic kineties of Strobilidium caudatum form a spiral at the posterior pole. Strobilidiids without such a spiral are transferred to the genus Rimostrombidium. Fourteen new combinations and a nomen novum, Strobilidium kahli, are necessary, Meseres corlissi n. sp. is characterized by eight somatic kineties composed of long cilia which are not fused to “bristles” as they are in Halteria. Strombidium oblongum shows similar characteristics and is thus combined with Meseres. Strombidium rehwaldi n. sp. has an anterior and an equatorial girdle of extrusomes. The morphogenesis of Meseres and Halteria is very similar, i.e. the entire somatic ciliature and the oral primordium originate apokinetally on the cell surface; the parental somatic ciliature is resorbed. In strobilidiids and tintinnids, the oral anlagen develop in a subsurface pouch and the parental somatic kineties, which are not resorbed, elongate by intrakinetal proliferation of basal bodies. In strombidiids, the oral primordium develops in an intracellular sac or tube. These morphogenetic peculiarities and distinct morphologic characters (e.g. arrangement of adoral membranelles) were applied in constructing a phylogenetic system for oligotrichs using hypotrichs as outgroup. This shows that halteriids are more closely related to hypotrichs than they are to other oligotrichs. The Halteriidae are thus raised to ordinal and subclass ranks, Halteriida n. ord., Halteriia n. subcl.

A MODIFICATION FOR PARAFFIN SECTIONS OF SILVER CARBONATE IMPREGNATION FOR MICROGLIA.

A technique is described whereby the cytoplasm of microglia is stained in routinely fixed paraffin embedded sections, greatly facilitating the study of these cells in normal and pathological sites. Of interest is the apparent paucity of these cells in normal tissues and their rapid and intensive proliferation in pathological states. The changes observed may be correlated with changes recognized with other stains on companion paraffin sections of the same tissue.

OLIGODENDROGLIOMAS

THE RECOGNITION of the oligodendrocyte as a cellular unit of the neuroglial portion of the central nervous system was made by Ford Robertson, a Scottish investigator, in 1900. With the use of platinum impregnation he separated a cell type from the third element of Cajal, which he called "mesoglia." Hortega, in 1918, analyzed the third element of Cajal with the use of silver carbonate impregnation methods and discovered first the microglial cell, which he called "microglia"; in 1921 he identified the oligodendroglial cell, which he so designated because of its few dendritic processes. The mesoglial cell of Robertson was found to correspond to the oligodendroglial cell of Hortega, but the term used by the latter was retained, since it was more descriptive and avoided implication of mesodermal derivation. Bailey and Cushing, 1 in their historic classification of gliomas, published in 1926, were the first to associate the oligodendroglial cell with