Reptiles collected in the vicinity of the Iowa Lakeside Laboratory, Dickinson County, Iowa, during the summer of 1950, were examined for helminths by Burnham (1951) and for protozoa by Schmidt (1954). lvlonocercomonas colubrorum, Trichomonas batrachorum and Haemogregarina sp. were found in both Thamnophis radix and T. ordinatus parietalis. An enteric flagellate of the genus Trimitus was found in a single specimen of T. radix. Special consideration is herein given to Trimitus because though this genus has been reported from North American amphibians and reptiles it appears that neither descriptions nor figures have been published, and also, because there is some uncertainty concerning the characterization of the genus

Saxe, L. H.

Schmidt, Eugene M.

English

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Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snake, Thamnophis radix

University of Northern Iowa

Proceedings of the Iowa Academy of ScienceVolume 60 | Annual Issue Article 991953Trimitus parvus Grasse (Protozoa, Mastigophora)from a garter snake, Thamnophis radixL. H. SaxeState University of IowaEugene M. SchmidtState University of IowaCopyright © Copyright 1953 by the Iowa Academy of Science, Inc.Follow this and additional works at: https://scholarworks.uni.edu/piasThis Research is brought to you for free and open access by UNI ScholarWorks. It has been accepted for inclusion in Proceedings of the Iowa Academyof Science by an authorized editor of UNI ScholarWorks. For more information, please contact scholarworks@uni.edu.Recommended CitationSaxe, L. H. and Schmidt, Eugene M. (1953) "Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snake, Thamnophisradix," Proceedings of the Iowa Academy of Science: Vol. 60: No. 1 , Article 99.Available at: https://scholarworks.uni.edu/pias/vol60/iss1/99Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snake, Thamnophis radix By L. H. SAXE AND EUGENE M. SCHMIDT INTRODUCTION Reptiles collected in the vicinity of the Iowa Lakeside Labora-tory, Dickinson County, Iowa, during the summer of 1950, were examined for helminths by Burnham ( 1951) and for protozoa by Schmidt ( 1954). lvlonocercomonas colubrorum, Trichomonas ba-trachorum and H aemogregarina sp. were found in both Thamn-ophis radix and T. ordinatus parietalis. An enteric flagellate of the genus Trimitus was found in a single specimen of T. radix. Special consideration is herein given to Trimitus because though this genus has been reported from North American amphibians and reptiles it appears that neither descriptions nor figures have been published, and also, because there is some uncertainty con-cerning the characterization of the genus. METHODS For the study of enteric protozoa smears of gut contents were fixed in Schaudinn's fluid with 5% glacial acetic acid added and either stained with Heidenhain's hematoxylin or impregnated by the protein-silver method of Moskowitz ( 1950). OBSERVATIONS Trimitus from T. radix is ovoid, with the anterior end usually somewhat broader than the posterior end. The size ranges from 4 to 6 microns in length and from 3 to 5 microns in width. [ Figures 1-3. Trimitus parvus from Thamnophis radix, fixed in Schaudinn's fluid with 5% glacial acetic acid added and impregnated by the protein-silver method of Mosko-witz. Figures were drawn by Schmidt with the aid of a camera lucida at an initial magnification of 4,000. 754 1Saxe and Schmidt: Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snaPublished by UNI ScholarWorks, 19531953] TRIMITUS PARVUS GRASSE 755 The vesicular nucleus is spheroidal in shape and located at the extreme anterior end of the body (see figures). The endosome is large and usually in contact with the nuclear membrane in the region at which the nuclear membrane is contiguous with the body surface. One or two dense granules are commonly seen within the endosome. The periendosomal space appears to be devoid of granules and may be an artifact of fixation. There are two anterior flagella of unequal length, that have an origin near the nucleus. The longer of these two flagella is ap-proximately the length of the body or slightly longer. The shorter flagellum is generally less than the length of the body. In protein-silver material, the flagella appear to be of uniform diameter, ex-cept for a proximal portion which is about one micron in length and about twice as thick as the rest of the filament. Also originating in the anterior of the body is the posterior fla-gellum, which has an enlarged proximal portion similar to that of the anterior flagella. The posterior flagellum is approximately twice the length of the body and is appressed to the body surface. Originating near the nucleus, along with the flagella, is a chro-matic rod-like structure, which extends posteriorly in the cytosome. It may be in juxtaposition with the posterior flagellum or indepen-dent of it. In either case, it does not protrude through the cytosom-al surface. It is analogous, if not homologous, to the structure in Enteromonas that Nie (1950) called the funis. The cytoplasm is vacuolated, with endocytoplasmic granules oc-casionally present. The authors have been unable to distinguish a blepharoplast with certainty. In silver-protein preparations the structure probably does not impregnate (see figure 3) ; in Heidenhain's hematoxylin material, it is obscured by a chromaticity of the cytoplasm surround-ing the nucleus. In their Trimitus material the authors have failed to find either a parabasal body or an axostyle. DISCUSSION Alexeieff ( 1910) found, in the digestive tract of a marme fish, Onos (= M otella) tricirrata, a small flagellate for which he pro-posed the name Trimitus motellae. He described the organism as being 6 to 8 microns in length; ovoid and slightly flattened in form with the posterior end drawn out and the anterior end often trun-cated. The mid-region of the body was filled with ingested bac-teria and surrounded by a layer of more dense peripheral cytoplasm. The nucleus was found in the extreme anterior of the body. It was, said Alexeieff, the flagella which characterized this organism. There 2Proceedings of the Iowa Academy of Science, Vol. 60 [1953], No. 1, Art. 99https://scholarworks.uni.edu/pias/vol60/iss1/99756 IOWA ACADEMY OF SCIENCE [Vol. 60 were two anterior flagella of unequal length (the one shorter, the other a bit longer than the body) and also a long trailing flagellum (up to four times the body length) that was joined, in its proximal portion, to the body surface. Duboscq and Grasse ( 1923) described flagellates from Kalo-termes which they placed in the genus Trimitus. Kirby ( 1930) erected the genus Tricercomitus to include these organisms and de Mello ( 1932) also classified Trim it us-like flagellates from a termite as Tricercomitus. Among differences which distinguish Tricerco-mitus Kirby from Trimitus Alexeieff are the presence in the former of three equal anterior flagella, a parabasal body and an axostyle. Grasse ( 1952), however, considered Tricercomitus to be a synonym of Trimitus. Grasse ( 1932), from Rana tem poraria and Buf o vulgaris, de-scribed Trimitus parvus with a trailing posterior flagellum about twice the body length and two anterior flagella, one shorter, the other the same length or longer than the body. Many individuals were described to have a third anterior flagellum. A rod shaped parabasal body and a lightly staining filiform axostyle were also described. Wenrich in the course of culture experiments has identified Trimitus parvus from Rana pipiens (1945, 1947) and Triturus viri-descens ( 1947), and Trimitus sp. from Pseudemys floridana mobil-ensis (1947). Trimitus was previously reported from a turtle by Knowles and Das Gupta ( 1930) . Wenrich and Saxe (unpublished) found Trimitus in a specimen of Thamnophis sirtalis captured in Delaware County, Pennsylvania. Moskowitz (personal communication), in his study of the flagellates of Squamata ( 1951), recognized but did not record the presence of Trimitus in his material. In the present study the authors confirm the generic character-ization given by Alexeieff (1910). No structure identifiable as a parabasal body was found even in material impregnated by a pro-tein-silver technique. Silver impregnation procedures are widely recognized as the most satisfactory of methods for the demonstra-tion of Janicki-type parabasal bodies and the technique employed successfully revealed the parabasal apparatus of Trichomonas batra-chorum, present along with Trimitus. Also, the authors have failed to distinguish an axostyle in Trimitus from either Thamnophis or Rana. In the present material, Trimitus with more than two anterior flagella were found only when division was obviously in progress. 3Saxe and Schmidt: Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snaPublished by UNI ScholarWorks, 19531953] TRIMITUS PARVUS GRASSE 757 In examining material from Rana pipiens an occasional specimen of M onocercomonoides, possibly M. rotunda (Bishop), was en-countered. This finding suggests that some "Trimitus" with more than two anterior flagella may have been mistakenly identified. In this connection the resemblance of Grasse' s figures ( 195 2) of Tri-m it us blattae to Monocercomonoides orthopterorum is striking. The presence of a funis in Trimitus is considered evidence of a relationship to Enteromonas. These two genera differ significantly only in the number of anterior flagella, two in Trimitus, three in Enteromonas. If Trimitus truly lacks an axostyle and parabasal body, which the present investigation has not revealed, the genus must be removed from the Trichomonadida where 1t is placed by Grasse ( 1952) and join Enteromonas, possibly in the Cercomonad-idae as suggested by Kirby ( 1930) . A comparison of flagellates of the genus Trimitus from Thamn-ophis radix and Rana pipiens has shown that these organisms are of the same species, Trimitus parvus. The snake might well have acquired its infection by feeding upon frogs, as larger garter snakes commonly do. SUMMARY 1. Observations have been made that confirm Alexeieff's charac-terization of the genus Trimitus. 2. The same species, Trimitus parvis, may occur in both Thamn-ophis radix and Rana pipiens and the former host may acquire its infection from its prey. 3. Trimitus is closely related to Enteromonas and does not be-long in the order Trichomonadida. Literature Cited Alexeieff, A. 1910 Sur les flagelles intestinaux des poissons marins (Note preliminaire). Arch. Zool. Exper. et Gen. Series 5. 6(Notes et Rev.): I-XX. Burnham, K. D. 1951 Helminths in terrestrial reptiles of north-western Iowa, M. S. Thesis, State University of Iowa. Duboscq, 0. and P. P. Grasse 1923 Sur les petits flagelles de Calotermes flavicollis ( Fab.) Com pt. Rend. A cad. Sc., Paris. 177 :362-364. Grasse, P. P. 1932 Sur un nouveau flagelle parasite du genre Trimitus Alex. Com pt. Rend. Soc. Biol., Paris. lll :840-843. Grasse, P. P. 1952 Ordre des Trichomonadines. Chapter in Grasse (editor): Traite de Zoologie Tome I, fasc. l. Kirby, H. 1930 Trichomonad flagellates from termites. I Tricercomitrts gen. nov. and Hexamastix Alexeieff. Univ. Calif. Publications Zool. 33:393-444. Knowles, R. and B. M. Das Gupta 1930 On two intestinal protozoa of an Indian turtle. Indian J. Med. Research 18 :97-104. de Mello, F. 1932 On the nature and identification of some small trichomonads from the intestine of termites hitherto related to the so-called Trimitus 4Proceedings of the Iowa Academy of Science, Vol. 60 [1953], No. 1, Art. 99https://scholarworks.uni.edu/pias/vol60/iss1/99758 IOWA ACADEMY OF SCIENCE [Vol. 60 stages of Duboscq and Grasse. (Abstract) Proc. Indian Sc. Cong. 19:259. (Cited from Biol. Abstr. 8 :9693). Moskowitz, N. 1950 The use of protein silver for staining protozoa. Stain Tech. 25:17-20. Moskowitz, N. 1951 Observations on some intestinal flagellates from reptilian host (Squamata). J. Morphol. 89:257-321. Nie, D. 1950 Morphology and taxonomy of the intestinal protozoa of the guinea pig, Cavia porcella. J. Morphol. 86:381-493. Schmidt, E. M. 1954 Observations on endozoic protozoa from two species of Thamnophis. M. S. Thesis, State University of Iowa. Wenrich, D. H. 1945 The cultivation of Trichomonas augusta (Protozoa) from frogs. J. Parasitol. 31 :375-380. Wenrich, D. H. 1947 Culture experiments on intestinal flagellates. III Species from amphibians and reptiles. J. Parasitol. 33 :62-70. IowA LAKESIDE LABORATORY MILFORD, IOWA DEPARTMENT OF ZOOLOGY STATE UNIVERSITY OF IOWA IowA CITY, IowA 5Saxe and Schmidt: Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snaPublished by UNI ScholarWorks, 1953

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Trimitus parvus Grasse (Protozoa, Mastigophora) from a garter snake, Thamnophis radix

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