Acanthamoeba
Acanthamoeba Volkonsky, 1931 (ref. ID; 36877, 7298, 7358, 7710) reported year? (ref. ID; 1618)
Phylum Rhizopoda Von Siebold, 1845: Class Lobosa Carpenter, 1861: Order Acanthopodina Page, 1976: Family Acanthamoebidae Sawyer & Griffin, 1977 (ref. ID; 7298, 7358)
Order Acanthopodida Page, 1976 (ref. ID; 6789)
[ref. ID; 177]
Members of the genus Acanthamoeba are ubiquitous freshwater and soil amoebae that occur world-wide. During the last decade (1982~), these amebae have been isolated with increasing frequency from contact lens paraphernalia as well as from human tissue such as corneal scrapings and the central nervous system (Ma et al. 1990; Visvesvara & Stehr-Green 1990). Acanthamoeba can be readily recognized because of the striking morphologic characteristics of the trophozoites and cysts. Based on the size and morphologic features of cysts, Pussard & Pons (1977) established 18 species and placed of them in three groups. Group 1 now consists of four species (A. astronyxis, A. comandoni, A. echinulata, and A. tubiashi) characterized by large trophozoites and cysts (>/_ 18 um) (Page 1988; Visvesvara 1991). Species in groups 2 and 3 are smaller (< /_ 18 um) but can be separated from each other on the basis of cyst morphology. Group 2 comprises 10 species (A. castellanii, A. polyphaga, A. rhysodes, A. mauritaniensis, A. divionensis, A. griffini, A. lugdunensis, A. quina, A. hatchetti, and A. triangularis), and five species (A. culbertsoni, A. lenticulata, A. palestinensis, A. pustulosa, and A. royreba) are in group 3 (Page 1988; Visvesvara 1991). Although cyst characteristics make genus identification easy, differentiation to the species level is difficult. This is especially true for members of groups 2 and 3 which cannot always be identified by species on a morphologic basis alone. As a result, the most recent identification attempts have been based on both morphologic and biochemical criteria such as isoenzymes, antigens, protein profiles, and restriction fragment-length polymorphism analysis of DNA resolved by electrophoretic methods (Byers et al. 1983; DeJonckheere 1983; Page 1988; Visvesvara 1991). Cladistic analysis of the data has also been used to elucidate the affinities of the various species of Acanthamoeba (Dagget et al. 1985). (ref. ID; 177)
[ref. ID; 1618]
Small amoebae similar to Hartmannella; ectoplasm is not well developed; mitotic figure at the end of metaphase, a straight or concave spindle with sharply pointed poles. Cysts enveloped by 2 membranes, the outer envelope being highly wrinkled and mammillated. Several species. Neff (1957) obtained axenic cultures of a species from soil. The presence of Acanthamoeba in tissue cultures of trypsinized monkey-kidney cells has been reported by several observers since 1957. (ref. ID; 1618)
[ref. ID; 3701]
The classification of the small soil and freshwater amoebae, particularly those assigned to the genus Acanthamoeba, has been based entirely on the organisation of the nucleus (Singh 1951, 1952; Singh & Das 1970) or on the structure of the trophozoite or cyst (Page 1967, 1976). Comparisons of a wide range of strains using immunological (Willaert 1976), genetic (Adam & Blewett 1974) and physiological techniques (Adam 1064; Griffiths, Curnick, Unitta & Wilcox 1978) have, however, indicated that morphological criteria are not adequate for estimating taxonomic relationships between these organisms. (ref. ID; 3701)
[ref. ID; 3991]
Type species; Acanthamoeba castellanii (Douglas, 1930) (ref. ID; 3991)
[ref. ID; 4022]
Trophozoites of the different species of Acanthamoeba are difficult to identify because of the absence of distinctive morphologic features. The shape and the structure of cysts, on the other hand, differ considerably among the species and can be used as valid taxonomic criteria. (ref. ID; 4022)
[ref. ID; 7430]
Acanthamoeba has been isolated from air, soil, freshwater, saltwater, wild animals and humans. Identification of amoebae from the genus Acanthamoeba is relatively easy after they have been grown in culture. Classification at the subgenus level is a problem, however, even for expert taxonomists. (Visvesvara 1991). The nomenclature and taxonomy of Acanthamoeba have been revised many times (Visvesvara 1991), most recently ~20 years ago (Pussard & Pons 1977; Sawyer & Griffin 1975). Pussard and Pons divided the genus into three morphological groups based on cyst size and shape. Group 1 species have large cysts compared to the other groups, with a smooth ectocyst and stellate endocyst. Group 2 species have a wrinkled ectocyst and the endocyst can be stellate, polygonal, triangular, or oval. Group 3 species typically have a thin, small ectocyst with a round endocyst. However, Sawyer first observed that ionic strength of the growth medium can alter the shape of cysts walls (Sawyer 1971), thus, substantially reducing the reliability of cyst morphology as a taxonomic charateristic. Pussard and Pods (1977) used the number of ostioles in the cysts as defining characters, but this character has been mentioned infrequently in subsequent descriptions of species. In the last decade, several groups have used analysis of isoenzyme electrophoretic patterns to address intrageneric relatioships and to test the morphological classification scheme. These studies discoverd extensive diversity among isolates of Acanthamoeba as exhibited by unique isoenzyme patterns, especially for group 1 species. Moura, Wallace and Visvesvara (1992) found good agreement between isoenzyme patterns and morphological groups, but their study was limited and included only one group 2 isolate, A. castellanii Castellani. De Jonckheere (1983), Daggett et al. (1983), and Costas and Griffiths (1986), studied larger groups of isolate including 30, 71 and 37 strains, respectively. Each study divided isolated of Acanthamoeba into several different groups that often were inconsistent with species and/or morphological group designations. Each study suggested a need for revision of the classification. Analyses of Acanthamoeba mitochondrial DNA restriction fragment length polymorphisms (mtRFLP), although limited in number of isolates used, also revealed a large degree of interstrain genetic diversity (Bogler et al. 1983; Yagita & Endo 1990). Species belonging to groups 2 and 3, especially A. castellanii and A. polyphaga, were shown to be polyphyletic.
Isoenzyme patterns and mtRFLP analyses have highlighted ambiguities in the morphology-based classification scheme, but neither approach directly addressed the phylogeny of Acanthamoeba. Johnson et al. (1990) analyzed partial nuclear 18S rRNA sequences from seven isolated of Acanthamoeba and obtained results that were concordant with the classification of Pussard and Pons, but only five species were included. Our lab recently began an investigation of 18S rRNA gene (Rns) phylogeny using 18 isolates of Acanthamoeba from morphological groups 2 and 3 (Gast et al. 1996). (ref. ID; 7430)
[ref. ID; 7714]
The classification of Acanthamoeba at the species level is unsatisfactory in that there are poor correlations between morphology and physiological (Adam 1964; Griffiths, Curnick, Unitt and Wilcox 1978), genetic (Adam and Blewett 1974), immunological (Visvesvara and Balamuth 1975; Willaert and Jadin 1974) and biochemical (Costas and Griffiths 1980) characters. The fatty acid profiles (22 fatty acid methyl acid methyl esters) of the strains varied from batch to batch and at different growth temperatures to such an extent that it was not possible to distinguish between the species. (ref. ID; 7714)
- Acanthamoeba astronyxis (Ray & Hayes, 1954) (ref. ID; 299) reported year? (ref. ID; 1543) reported author and year? (ref. ID; 177, 3701, 3985, 4181, 4218, 7430)
- Acanthamoeba castellanii Douglas, 1930 (ref. ID; 3687) reported year? (ref. ID; 1543, 1618, 3847) or (Douglas) Volkonsky (ref. ID; 1335) reported author and year? (ref. ID; 177, 3701, 4110, 4158, 4173, 4181, 4218, 4325, 4694, 6791, 7430)
Syn; Hartmanella rhysodes Singh, 1952 (ref. ID; 4694) reported year? (ref. ID; 4158, 4173)
- Acanthamoeba comandoni (ref. ID; 177, 3793, 7430)
- Acanthamoeba culbertsoni (Singh & Das) (ref. ID; 3847) reported author and year? (ref. ID; 177, 3639, 4110, 7430)
- Acanthamoeba divionensis (ref. ID; 177, 4110)
- Acanthamoeba echinulata (ref. ID; 177)
- Acanthamoeba griffini Sawyer, 1971 (ref. ID; 7358) reported author and year? (ref. ID; 177, 4110, 4158, 4181, 4218, 4694, 7430, 7661)
- Acanthamoeba hatchetti Sawyer (ref. ID; 3847) reported author and year? (ref. ID; 177, 4110, 4181, 7430)
- Acanthamoeba healyi Moura, Wallace & Visvesvara, 1992 (ref. ID; 177 original paper, 7430)
- Acanthamoeba hyalina Dobell & O'Conner, 1921 (ref. ID; 3687) reported year? (ref. ID; 1618)
- Acanthamoeba jacobsi Sawyer et al., 1992 (ref. ID; 7358)
- Acanthamoeba lenticulata (ref. ID; 177, 4110, 7430)
- Acanthamoeba lugdunensis (ref. ID; 177, 4110, 7430)
- Acanthamoeba mauritaniensis (ref. ID; 177, 4110)
- Acanthamoeba palestinensis (Reich, 1933) (ref. ID; 4090) or (Reich, 1933) Page, 1967 (ref. ID; 3767) reported author and year? (ref. ID; 177, 3701, 4022, 4110, 4181, 4218, 7430)
Syn; Mayorella palestinensis Reich, 1933 (ref. ID; 3767)
- Acanthamoeba paradivionensis (ref. ID; 4110)
- Acanthamoeba pearcei Nerad et al., 1995 (ref. ID; 7358 original paper) reported author and year? (ref. ID; 7430)
- Acanthamoeba polyphaga Puschkarew, 1913 (ref. ID; 4105) reported year? (ref. ID; 1543, 3847) reported author and year? (ref. ID; 177, 3639, 3701, 4110, 4158, 4181, 4218, 4258, 4694, 7430)
- Acanthamoeba pustulosa (ref. ID; 177, 4110, 7430)
- Acanthamoeba quina (ref. ID; 177, 4110)
- Acanthamoeba rhysodes (Singh, 1952) (ref. ID; 4269) reported year? (ref. ID; 3847) reported author and year? (ref. ID; 177, 3639, 3701, 4110, 4181, 4218, 7430)
- Acanthamoeba royreba Willaert, Stevens & Tyndall, 1978 (ref. ID; 3805 original paper, 4110) reported author and year? (ref. ID; 177)
- Acanthamoeba stevensoni Sawyer, Nerad, Lewis & Mclaughlin, 1993 (ref. ID; 7298 original paper, 7358, 7430)
- Acanthamoeba terricola (ref. ID; 4181, 4218)
- Acanthamoeba triangularis (ref. ID; 177)
- Acanthamoeba tubiashi (ref. ID; 177, 7430)
Acanthamoeba astronyxis (Ray & Hayes, 1954) (ref. ID; 299) reported year? (ref. ID; 1543) reported author and year? (ref. ID; 177, 3701, 3985, 4181, 4218, 7430)
Descriptions
- Trophozoite: The rough endoplasmic reticulum (RER) is abundant and appears as short cisternae scattered randomly throughout the cytoplasm. The nucleus contains one large nucleolus, within which a clear centrally located space is usually seen. Occasionally the nucleolus appears ring-shaped. A structure is found beside the nucleus that corresponds to the centrosphere described by light microscopists in living and stained specimens of interphase and dividing cells. The centrosphere consists of Golgi complex and an electron dense body from which microtubules radiate. The Golgi complex is composed of several stacks of elongated cisternae and numerous small vesicles. The electron-dense body appears either as a small amorphous, dense organelle or as a cylindrical structure with a length of ca. 500 nm and ca. 150 nm wide with a moderately electron-dense, narrow lumen and very electron-dense walls. This cylindrical structure, noted only in longitudinal sections, resembles the centriole in appearance and size. This organelle has never been observed in cross section, making it impossible to determine if a triplet microtubule structure exists. In addition to that in the centrosphere, numerous other well developed Golgi complexes are found elsewhere in the cytoplasm. Mitochondrial profiles are numerous and are either spherical, ovoid, or dumbbell-shaped. They usually contain a grainy electron-dense vesicle. Relatively few lipid droplets, which have different diameters, are visible in the cytoplasm. Lipid droplets and mitochondrial profiles are ordinarily to interspersed among each other. (ref. ID; 3985)
- Encysting cells: The first apparent change observed at the onset of encystment is the reorganization of the RER. The cisternae appear elongated and curved and are found in closer proximity to the various organelles. As encystment progresses, circles of RER appear, enclosing various protoplasmic inclusions, especially mitochondria. Occasionally, cisternae stacked in parallel arrays are located near the periphery of the cell. Concomitantly, mitochondrial profiles show a marked tendency toward elongation and constriction. Most prominent are curved mitochondria, which often have a shape of an open ring. The ends of these mitochondria appear normal, while the curved mid-portion consists of closely apposed longitudinally-oriented membranes. Some mitochondria are dumbbell-shaped, horseshoe-shaped, or annular. During encystment protrusions from nucleoli are noted, which appear to pinch off as nucleolus-like bodies. One or two such bodies per nucleus are observed. These structures are spherical with the same electron densities as the nucleoli. Some nucleoli have a large central vacuole surrounded by a ring of electron-dense material. In a later stage of encystment, the cell becomes round. The cytoplasm becomes more and more dense. Mitochondria decrease in size and numbers. The amount and concentration of lipid droplets increase considerably. The final deposition of an outer wall, the exocyst, around the cell is indicative of the formation of the young cysts. (ref. ID; 3985)
- Young cyst: The exocyst is very close to the cell membrane and appears as a layer of granular material ca. 200 nm in thickness. The cytoplasm is filled with numerous lipid droplets, autolysosomes, and small, usually spherical, mitochondria. The RER often assumes the form of whorls a consisting of 2-8 concentric layers of profiles, surrounding the mitochondria, nucleus, food vacuoles, or other inclusions. Long cisternae arranged in rows parallel to the cell surface are often observed. The young cyst is transformed into a mature cyst upon completion of the deposition of an inner wall, the endocysts, between the cell membrane and the exocyst. (ref. ID; 3985)
- Mature cyst: The mature cyst is surrounded by an almost circular exocyst and a star-shaped endocyst that is closely apposed to the cell membrane. The exocyst is separated from the endocyst by a clear zone of variable width, which contains fragments of membranes or membrane-bounded vesicles. The endocyst (~300-400 nm) is thicker than the exocyst and contains larger granules. In the region of the rays. The endocyst is elevated and fuses with the exocyst, thus forming a bow-shaped space, the ostiole. The ostiole is occupied by a large bell-shaped structure, the operculum, that lies close to the underlying plasma membrane on its concave surface. The operculum is composed of granular material, similar to that of the endocyst. The ostiole is limited above the operculum by a very thin layer of the exocyst, which is continuous over the entire surface of the cyst. An excysting amoebae exists through the ostiole after displacement of the operculum. The cytoplasm of the mature cysts contains small mitochondria, autolysosomes, and many lipid droplets. The lipid droplets occur singly or as aggregates. Abundant RER is observed mainly in those sections cut through the plane of nucleus. Although whorls of the RER are absent, circles of RER enclosing mitochondria are still found in mature cyst. (ref. ID; 3985)
Acanthamoeba castellanii Douglas, 1930 (ref. ID; 3687) reported year? (ref. ID; 1543, 1618, 3847) or (Douglas) Volkonsky (ref. ID; 1335) reported author and year? (ref. ID; 177, 3701, 4110, 4158, 4173, 4181, 4218, 4325, 4694, 6791, 7430)
Synonym
Hartmanella rhysodes Singh, 1952 (ref. ID; 4694) reported year? (ref. ID; 4158, 4173)
Descriptions
In association with fungi and certain bacteria; Hewitt obtained the organism from agar cultures of sample soil taken from among the roots of white clover; co-existing with yeast-like fungi, Flavobacterium trifolium and Rhizobium sp.; some cysts are said to remain viable at 37 degrees C for 6 days. (ref. ID; 1618)
Marine amoeba. (ref. ID; 3847)
Acanthamoeba castellanii is a free-living organisms, found in soil and fresh water, which forms a dormant, non-reproductive cysts. The cyst has a thick, wrinkled outer wall and a cellulose-containing inner wall. The outer wall is resistant to chemical and physical analysis so that little is known about its composition and molecular organization. (ref. ID; 4173)
Measurements
12-30 um in diameter. (ref. ID; 1618)
Available culture
Acanthamoeba castellanii Neff strain (ATCC 30010). Genome sequence. (ref. ID; 6791)
Acanthamoeba hatchetti Sawyer (ref. ID; 3847) reported author and year? (ref. ID; 177, 4110, 4181, 7430)
Descriptions
Marine amoebae. (ref. ID; 3847)
Acanthamoeba hyalina Dobell & O'Conner, 1921 (ref. ID; 3687) reported year? (ref. ID; 1618)
Descriptions
According to Volkonsky, the organism described by Dobell & O'Conner as Hartmannella hyalina, is transferred to this genus. Small amoeba; a single contractile vacuole; binary fission; mitotic figure a sharply pointed spindle. Cysts spherical; with a smooth inner and a much wrinkled outer wall; easily cultivated from old faeces of man and animals; also in soil and fresh water. (ref. ID; 1618)
Measurements
9-17 um in diameter when rounded; cyst 10-15 um in diameter. (ref. ID; 1618)
Acanthamoeba palestinensis (Reich, 1933) (ref. ID; 4090) or (Reich, 1933) Page, 1967 (ref. ID; 3767) reported author and year? (ref. ID; 177, 3701, 4022, 4110, 4181, 4218, 7430)
Synonym
Mayorella palestinensis Reich, 1933 (ref. ID; 3767)
Descriptions
- Light microscopy: The cysts examined in an interference microscope appears as a spherical double-walled structure containing a central nucleus and peripherally situated granules of different size. The nucleus includes a large nucleolus and spherical nucleolus-like bodies of varying size and number; most frequently there are 3-5 such bodies. These structures have never been seen in trophozoites. The nucleolus-like bodies stain bluish-black with methyl green-pyronin, this staining not being changed appreciably after digestion with RNase. Feulgen reaction gave negative results. (ref. ID; 4022)
- Electron microscopy: Cyst wall; The cyst is nearly round, having only slight angles, and is covered by 2 distinct walls, an inner endocyst and an outer exocyst. The endocyst follows closely the shape of the protoplast and lies near the cell surface. The exocyst has an irregular shape and is, in most places, separated from the endocyst by a wide space filled in some regions with a spongy network or with cytoplasmic debris. The 2 walls hardly differ in thickness, texture, and staining characteristics. They appear as an optically dense granular layer 80-150 nm thick. The inner and the outer walls fuse at irregular intervals, forming gaps in the cyst wall that contain either endocyst nor exocyst. This ostiole-like region is occupied by a flat structure, the operculum, which lies closer to the plasma membrane than the endocyst. It has the same optimal appearance as the cyst wall. The number of ostioles varies from 1 to 3. No apparent differences have been found between mature cysts formed in old cultures and induced ones. (ref. ID; 4022)
- Nucleus. The interphase nucleus is typical containing 1 large nucleolus with electron-dense material. During encystment, changes in the nuclear envelope and the appearance of nucleus-like bodies have been observed. The nuclear membrane forms bulb-shaped projections. Vesicles enclosed by a double membrane identical with that of the nucleus have been found in the cytoplasm near the nucleus. These vesicles are probably derived from the membrane projections. The nucleolus-like bodies have a regular spherical shape and are more optically dense that the nucleolus. They consist of an amorphous condensed material and are surrounded by particles of the same appearance as those of the nucleolus. (ref. ID; 4022)
- Golgi complex. The Golgi complex of A. palestinensis is distinguished by its small size and compact cisternae. Several complexes are distributed throughout the cytoplasm but are most abundant in the perinuclear region. Each Golgi complex comprises two or more stacks of 3-6 smooth cisternae. The cisternae are usually curved to a greater or lesser extent, although straight flat cisternae are also observed. Cisternae are separated by a space of about 16 nm; the space between the membranes of a single cisternae is also approximately 16 nm, except at the margins where the sacs are slightly dilated. Many smooth, small vesicles and randomly oriented tubules are found at the periphery of the stacks or concentrated on the concave face of the stacked cisternae. The vesicles are about 50 nm in diameter and appear to arise from the ends of the cisternae. The relative proportion of the vesicular and cisternal components vary from stack to stack. Usually two Golgi complexes are found on the opposite sides of the nucleus. One of them often appears to be in an intimate association with the nuclear membrane. Occasionally a third complex is observed close to the nucleus. The nuclear membrane appears discontinuous in the region of the concave face of a Golgi complex. On one side of this discontinuity a small vesicle is noted. The other end of the membrane is clearly associated with a vesiculated end of a tubule which seems to be continuous with a cisternal membrane of the Golgi apparatus. Golgi-like vesicles are also seen to be associated with nuclear membrane. A relatively large Golgi complex is observed in an invagination of the nucleus. This Golgi complex consists of a stack of a few flat cisternae, tubules, and numerous vesicles. In the center a cylindrical structure from which microtubules radiate can be seen. The convex face of the Golgi complex is apposed to the nuclear membrane. The region of contact contains compact cisternae, vesicles of various sizes, as well as granular and amorphous electron-dense material. Structural changes in the nuclear envelope are also observed in this area. The membrane loses its characteristic sharp definition and fuzzy layer obscures it. In the same section a second Golgi complex is seen on the opposite side of the nucleus at some distance from it. This Golgi complex consists of two stacks of cisternae, one being curved and the other flat. Vesicles and tubules are found at the periphery of the cisternae. Microtubules are observed at the concave face of the curved cisternae and alongside the stack of flat cisternae toward the nucleus. Cisternae of rough endoplasmic reticulum (RER) are often seen to approach the Golgi apparatus closely. In some instances a cisterna of RER seems to be continuous with a Golgi cisterna. (ref. ID; 4090)
- Microbodies. In the cytoplasmic of A. palestinensis, structures resembling mitochondria in shape and size have been observed. These organelles however are surrounded by a single unit membrane and contain a granular matrix which is less dense than that a of mitochondria. Tubular inclusions are presented in the matrix. These tubules have a less-defined and narrower lumen than the tubular cristae of the mitochondria. The circular or ovoid contour of these bodies is irregular. The diameter of the circular bodies ranges from 0.7 um to 1.20 um. The dimensions of the ovoid structures are approximately 1.3 um in length and about 0.5 um in width. Usually a RER cisterna is found in association with the microbody's membrane. (ref. ID; 4090)
- Mitochondria. Mitochondrial profiles are abundant and appear in various shapes, spherical (~0.5 um in diameter), ovoid (~1x0.6 um) or elongated (~1.7x0.7 um). As in other amoebae, they contain intracristal granular inclusions. The matrix is dense and frequently contains one large, very dense droplet or irregular granule. Occasionally, mitochondria are found which are distinguished by the presence of two apposed double membranes, running through the midline of the mitochondrion. These membranes seem to be continuous with the inner mitochondrial membrane at one end of the mitochondrion but form blind enlargements at the opposite end. (ref. ID; 4090)
Comments
The inner and outer walls of the cyst of A. palestinensis have almost the same fine structure, while in A. castellanii they differ considerably (Bowers & Korn 1969). The endocyst of A. palestinensis is nearly rounded, the few ostioles are located anywhere, and the operculum is flat and close to the protoplast surface. In A. castellanii, on the other hand, the endocyst is star-shaped with several truncated rays, the ostioles are located at the edges of the rays, and the operculum is thick, arched and further away from the cell surface. (ref. ID; 4022)
Acanthamoeba pearcei Nerad et al., 1995 (ref. ID; 7358 original paper) reported author and year? (ref. ID; 7430)
Diagnosis
Trophozoites typical of the genus Acanthamoeba Volkonsky, 1931. Locomotive forms 25.0-42.5 um long (average=3.1) by 17.5-25.0 um wide (average=22.1) and feeding forms approximately as wide as long. Amoebae contract in presence of light or heat from the microscope altering typical features of locomotive and feeding forms. Fixed and stained specimens approximately 22.5-39.0 um long (mean 27.9) by 12.5-17.5 um wide (average=14.7); nucleus 3.5-4.0 um (average=3.5). Living cysts 17.5-25.0 um in diameter (average=20.5). Ectocyst smooth without depressions at tips of arms or rays of the endocyst. Endocysts with 3-7 rays or arms, rarely 8-9, that are deeply scalloped with blunt or rounded tips. Acanthamoeba pearcei, strain ATCC 50435 did not grow at 37-39 degrees C, and was not pathogenic to laboratoy mice via the intranasal route. (ref. ID; 7358)
Isoenzyme profiles
Zymograms for propionyl esterase (PE), leucine aminopeptidase (LA) and acid phosphatase (AP) by enzyme electrophoresis were distinct for A. pearcei (ATCC 50435) and morphologically similar A. astronyxis (ATCC 30137). The PE zymogram for A. pearcei had seven bands compared to five bands for A. astronyxis with only one band shared by both species. The zymograms for LA had a single band with that of A. pearcei being more anodal than that of A. astronyxis. The AP zymogram for A. pearcei had one discrete anodal band and two cathodal bands in contrast to six anodal and two cathodal bands for A. astronyxis. The single anodal band for A. pearcei appeared to be shared with one of the two anodal bands for A. astronyxis. Twenty-one bands were expressed by the two strains, but only four (19%) were shared. (ref. ID; 7358)
Notes
There are several species of Acanthamoeba, that are not readily identifiable solely on the basis of their morphological features. Pussard and Pons (1977) proposed that each species be placed into one of three groups on the basis of cyst morphology in order to facilitate preliminary identification. Group 1 was proposed for species with distinctly scalloped endocysts, including A. astronyxis, A. comandoni, and A. echinulata. Acanthamoeba pearcei clearly belongs to Group 1, most closely resembling A. astronyxis. The principle characteristics for distinguishing between the two are the features of their cysts and their distinctly different zymograms. The new species, A. pearcei, is the fifth species originally isolated from marine or brackish waters. The first, A. griffini, was found in Long Island Sound, New York (Sawyer 1971), the second, A. hatchetti, from Baltimore Harbor, MD (Sawyer et al. 1977), the third, A. jacobsi, from the discontinued New York 12-Mile nearshore waste disposal site (Sawyer et al. 1992), and the fourth, A. stevensoni, from contaminated shellfish beds near Staten Island, New York (Sawyer et al. 1993). Acanthamoeba hatchetti and A. stevensoni were highly pathogenic to mice, and A. jacobsi mildly pathogenic. Acanthamoeba griffini and A. pearcei were not pathogenic under the conditions tested and neither grew when incubated at 39-40 degrees C. (ref. ID; 7358)
Type locality
Sewage contaminated ocean sediments from the northeast Atlantic Ocean, 65 km offshore from Maryland and Delaware, USA, approximately 38 degrres 19.6'N, 74 degrees 18.0'W. (ref. ID; 7358)
Type strain
Deposited at the American Type Culture Collection, Rockville, Maryland 20852, Accession no. ATCC 50435. (ref. ID; 7358)
Acanthamoeba polyphaga Puschkarew, 1913 (ref. ID; 4105) reported year? (ref. ID; 1543, 3847) reported author and year? (ref. ID; 177, 3639, 3701, 4110, 4158, 4181, 4218, 4258, 4694, 7430)
Descriptions
Marine amoebae. (ref. ID; 3847)
Acanthamoeba rhysodes (Singh, 1952) (ref. ID; 4269) reported year? (ref. ID; 3847) reported author and year? (ref. ID; 177, 3639, 3701, 4110, 4181, 4218, 7430)
Descriptions
Marine amoebae. (ref. ID; 3847)
Acanthamoeba royreba Willaert, Stevens & Tyndall, 1978 (ref. ID; 3805 original paper, 4110) reported author and year? (ref. ID; 177)
Descriptions
- Electron microscopy: Trophozoite- At the ultrastructural level, the Oak Ridge strain was found to be similar to other Acanthamoeba species. The plasma membrane had a characteristic trilaminar structure, but lacked a cell coat. The cell surface was highly irregular, with numerous filar pseudopodia (acanthopodia) filled with a finely fibrillar material. This material, which composed the major part of the ectoplasm, was resolved as 5-nm fibrils at high magnifications. Fibrils of a similar size, appearance, and location were described in Acanthamoeba castellanii; They were identified as actin. The amebae contained numerous large, irregularly shaped phagocytic vacuoles and small vesicles, some of which undoubtedly represented elements of the smooth endoplasmic reticulum (ER). The reticulum, however, could not be identified as a discrete entity in the cytoplasm of the Oak Ridge strain. The contractile vacuole could be differentiated from the phagosomes by its associated structures. In most sections, only vesicles (50-100 nm in diameter) were observed at the periphery of the contractile vacuole; these were usually confined to ~50% of its surface. Profiles of the contractile vacuole with an elaborate system of tubules were rarely seen. It appears, however, that the vesicles must fuse with the tubules, which then empty directly into the contractile vacuole. The lumina of the tubules, which were ~0.25 um thick, were filled with a diffuse, lightly staining material that was largely absent from the vesicles. The rough ER was not as abundant in the Oak Ridge strain as in other species of Acanthamoeba. Elements of the rough ER were mainly in the form of short tubules randomly scattered throughout the cytoplasm. The tubules contained fine fibrillar material of low electron density. Although long cisternae were observed occasionally at the periphery of the endoplasm, parallel stacks of cisternae were not seen. Such a configuration of the rough ER was observed, however, in encysting cells. Most ribosomes, many in helical or rosette configurations, were free in the cytoplasm of the trophozoite. The most conspicuous organelles in the cytoplasm were the mitochondria, which were either spherical (~0.5 um diameter) or ovoid in shape (1.1x0.5 um). Cup or dumbbell-shaped mitochondria were not observed in the Oak Ridge isolate. The cristae were organized in a system of highly branching tubules (34 um in diameter), some of which extended the width of the mitochondria. The mitochondrial matrix was extremely dense and lacked granules; however, granules with electron-dense peripheries and less electron-dense cores were present within inflated regions of the cristae of some mitochondria. These granules had an average diameter of 0.1 um. They have been seen in mitochondria of various species of Acanthamoeba and have been referred to as intracristal bodies or amorphous granules; as yet their function is unknown. The Oak Ridge amebae had several well differentiated Golgi complexes. Each complex was composed of a stack of 5-7 flattened, smooth-surfaced sacs, dilated at the ends. Many small membrane-bounded vesicles were associated with both the outer and inner faces, but these were devoid of internal material as were the cavities of the Golgi cisternae. Two apparently different structures were often in close association with the Golgi complexes, and occasionally within the same cell. One of these was an electron-dense body that probably corresponds to the centriolar satelites previously described for A. castellanii. No substructure was evident in this body, but longitudinal and cross sections of microtubules were always associated with it. The other structure, noted only in longitudinal sections, more closely resembled a centriole. The structure was a short cylindrical body that had a moderately electron-dense lumen and very electron-dense walls. The density of the body was similar in stained and unstained sections. Within the lumen, however, a central dark line was evident. This line extended the length of the structure in all sections in which the "centriole" was seen. The lumen and walls measured ~26.5 nm and 32 nm in width, respectively. The outer edges of the centriole walls were associated with filamentous material from which radiated 25-nm microtubules. The microtubules were sparse in the cytoplasm in regions other than those around the Golgi complexes. The nucleus was bordered by a typical nuclear envelope, the outer membrane of which was studded with ribosomes. The membranes were separated by a distance of 20 nm and were interrupted at frequent intervals by nuclear pores filled with electron-dense material. In tangential sections, the pores measured 80 nm in diameter and had a central dense granule. The nucleolus, with an average diameter of 1.5 um, had well defined fibrous and granular regions. Vesiculated nucleoli were not seen. The nucleoplasm was finely fibrillar and granular and contained patches of more densely stained material, which was probably condensed chromatin. (ref. ID; 3805)
- Cyst: The mature cyst of Acanthamoeba is distinguished by its fully formed endocyst and exocyst. Its cytoplasm is usually so dense that it is difficult to describe adequately the ultrastructure of many of the organelles. For this reason, with the exception of those of the cyst wall, the observation of the Oak Ridge strain were confined mainly to young cysts that had endocysts in various degrees of completion and fully formed exocysts. No attempt was made to follow sequential alterations of the cytoplasmic organelles during termination of growth and induction of encystment. The most notable structures in the cytoplasm of the young cysts, which were absent in the trophozite, were numerous large membrane-bounded inclusion, some reaching 1.2 um in diameter. These structures, which contained extremely electron-dense material surrounded by lightly staining granular areas, apparently were autolysosomes. At high magnification degenerative forms of mitochondria were seen in the densely stained regions along with other nondescript vesicles, presumably of cytoplasmic origin. The dimensions of the granules in the lightly stained areas were approximately the size of ribosomes. The autolysosomes were evident in mature cyst, but their contents were much denser than those of the corresponding organelles in the young cyst. The rough ER appeared to be more abundant in cysts than in trophozites which was undoubtedly due to the greatly reduced volume of the cyst cytoplasm. Usually, short segments of rough ER cisternae were observed to be randomly distributed throughout the cytoplasm. Occasionally, however, cisternae, stacked in parallel arrays, were located near the periphery of the cyst. The parallel configuration was not noted in mature cysts, but rough ER could be resolved within the dense cytoplasmic matrix. The Golgi complex was well developed in young cysts that appeared to have almost completed exocysts, but no apparent endocyst. The Golgi complex, however, became less evident in young cysts with endocysts in various stage of completion and could not be identified with certainly in mature cysts. The cisternae of the Golgi complex in the encysting cell were widely dispersed and were not arranged in the typical stacks. The cavities of the cisternae were filled with material of a density resembling that of the substance found in small membrane-bounded vesicles located in close proximity to the complex. Similar vesicles were dispersed throughout the cytoplasm and adjacent to the plasma membrane, especially in the region of the ostiole. Phagocytic vacuoles were typically not seen in young cysts, but the contractile vacuole with its subsidiaries was clearly evident in the densely stained cytoplasm. The contractile vacuole was smaller than in the trophozoite and was associated with several vacuoles of sizes intermediate between the major and subsidiary vacuoles. The tubular system associated with the contractile vacuole of the trophozoite was not found in cysts. The mitochondoria underwent the most striking alterations during encystment of the Oak Ridge strain. In contrast to the trophozoite, most of the mitochondria in the young cysts were spherical, had poorly defined cristae, and contained a single large electron-dense droplet that occupied a considerable volume of the intracristate space. This structure, with a diameter of 0.15 to 0.3 um, ranged from central to peripheral in its position. Frequently, images were seen which suggested that the droplets were being released from mitochondria; structures of a similar size and density were observed also to be free in the cytoplasm. Although identical droplets, may originate from the intracristal bodies of trophozoite mitochondria, their function is unknown. The nucleolus was considerably reduced in size in the young cysts. It averaged ~0.57 um in diameter and consisted mainly of granular material with few patches of fibrillar substance. The nucleolus is seen surrounded by a halo of granular material with a lower electron density than that of the surrounding nucleoplasm; however, this was to characteristic of young cysts. The size and structure of the nucleolus were essentially the same in the mature cyst, but in a small percentage of cells, 1 or 2 extremely electron-dense, spherical bodies, with a diameter of 0.5-1 um, were seen in close association with the nucleolus. Although no counts were made, there seemed to be a striking decrease in the numbers of nuclear pores during maturation of the cyst. In many sections of the mature cyst nucleus, no pores were observed. The exocyst of the Oak Ridge strain was ~0.42 um thick. It was separated from the endocyst by a clear zone of variable width. The exocyst was fibrillar, with the peripheral fibrils more densely packed than those in the major portion of the exocyst. The fibrils were oriented mainly in layers parallel to the cell surface, but in some regions of the exocyst, they were organized in a reticular fashion. Small vesicles, vacuoles of irregular shape, and amorphous material occasionally were embedded in the exocyst. Presumably these components represented cytoplasmic debris that was extruded and trapped in the wall material during its secretion. In contrast with the exocyst, the mature endocyst, ~0.13 um thick, was granular in texture. The granular material, however, was not uniformly dispersed throughout the endocyst, but was condensed to various degrees near the surface of the organism. The endocyst was separated from the underlying plasma membrane by a space of 20 nm. This area, of low electron density, was filled with amorphous material in which vesicles were sometimes observed. On the cytoplasmic side, a thin rim of densely staining material was closely applied to the plasma membrane. The exocyst and endocyst were continuous only in the region of the ostioles, which were numerous in the Oak Ridge cyst. The operculum was composed mainly of endocysts material, except at the outer edge, which consisted of the peripheral densely staining zone of the exocyst. The operculum appeared to be well developed even in an early stage of endocyst secretion. Often, numerous membrane-bounded vesicles, many of which resembled those associated with the Golgi complexes were closely associated with the plasma membrane in the ostiole region. The vesicles, which invariably were seen only in young cysts, ranged from 40 nm to 130 nm in diameter. Some were completely filled with electron-dense material, whereas others had clear to lightly staining cores that were surrounded by an electron-dense periphery. Membrane-bounded vesicles, largely devoid of internal contents, also were seen in spaces between the operculum and the plasma membrane. (ref. ID; 3805)
Acanthamoeba stevensoni Sawyer, Nerad, Lewis & Mclaughlin, 1993 (ref. ID; 7298 original paper, 7358, 7430)
Diagnosis
Trophozoites typical of the genus Acanthamoeba Volkonsky, 1931. Growth at 38-40 degrees C, and pathogenic to laboratory mice. Amoebae in locomotion often seen with a stretched adhesion uroid or collopodium with tufts of short spiny processes. Cysts irregularly rounded with polygonal or bluntly stellate endocyst. The ectocyst wall is single, refractile, slightly rippled, and not inflated or circular. Contour of the ectocyst similar to that of polygonal endocysts, irregularly rounded in stellate endocysts with blunt arms. (ref. ID; 7298)
Type locality
Sewage-contaminated sediments from hard clam beds (Mercenaria mercenaria), in Raritan Bay, New York near Staten Island, approximately 70 degrees 07'8" longitude and 40 degrees 30'05" latitude. (ref. ID; 7298)
Type specimens
Deposited at the American Type Culture Collection, Rockville, Maryland 20852, Accession No. ATCC 50388. (ref. ID; 7298)
Measurements
Length 27.5-42.5 um (mean 33.9) and width 15.0-20.0 long (mean 17.6) in the living condition. Fixed and stained amoebae 22.5-32.5 um long (mean 29.7) and 15.0-20.0 wide (mean 17.1). Nucleus 3.5-4.0 um in diameter. Living cysts 10.0-22.5 um (mean 16.5). (ref. ID; 7298)