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Cestoidea: Caryophyllidea) 126 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY No. 20-0160 B from the Research Foundation . 1968a. Vitellogenesis and eggshell for- of the State University of New York. mation in Caryophyllaeuf, laticeps (Pallas) and Canjophyllaeides Fennica (Schneider) Literature Cited (Cestodea: Caryophyllaeidea). Z. f. Para- Calentine, R. L. 1965. The biology and tax- sitenk. 30: 18-32. onomy of Biacetahulum (Cestoda: Caryo- . 1968b. Two new caryophyllaeid ces- phyllaeidae). J. Parasit. 51: 243-248. todes from the spotted sucker, Minytrema , and J. S. Mackiewicz. 1966. Mono- nielanops (Raf.) (Catostomidae). J. Parasit. bothrium ulmeri n. sp. (Cestoda: Caryophyl- 54: 808-813. laeidae) from North American Catostomidae. Self, J. T., and H. F. Timmons. 1955. The Tr. Am. Micr. Soc. 85: 516-520. parasites of the river carpsucker (Carpiodes Hunter, G. W., III. 1927. Notes on the carpio Raf.) in Lake Texoma. Tr. Am. Caryophyllaeidae of North America. J. Para- Micr. Soc. 74: 350-352. sit. 14: 16-26. Szidat, L. 1937. Archigetes R. Leuckart, 1878, Kennedy, C. R. 1965. Taxonomic studies on die progenetische Larve einer fiir Europa Archigetes Leuckart, 1878 (Cestoda: Caryo- neuen Caryophyllaeiden-Gattung Biacetabu- phyllaeidae). Parasitology 55: 439-451. Mackiewicz, J. S. 1964. Variations and host- lum Hunter, 1927. Zool. Anz. 119: 166-172. parasite relationships of caryophyllaeids (Ces- Timmons, H. F. 1951. A study of the hel- todea) from fish of Lake Texoma, Marshall minth parasites of the river carpsucker County, Oklahoma. J. Parasit. 50 (3, sec. 2): (Carpiodes carpio) in Lake Texoma. Master's 31 (abstract). thesis, University of Oklahoma, 49 p. The Chromosomes of Hunterella nodulosa Mackiewicz and McCrae, 1962 (Cestoidea: Caryophyllidea) JOHN S. MACKiEwicz1 AND ARTHUR W. JONES Department of Zoology and Entomology, University of Tennessee, Knoxville, Tennessee Cytological studies on tapeworms have getes, it is impossible to judge the worth of been almost entirely confined to Cyclophyllid- similar cytotaxonomic studies in the monozoic ean cestodes (for reviews see Jones 1945, tapeworms of the order Caryophyllidea. One Walton 1959). Exceptions have been the purpose of the present study, one of a pro- studies of four species of the Pseuclophyllidea jected series, is to learn if cytological informa- by Wolcott (1959), and Wikgren and Gus- tion on the Caryophyllidea will aid in the tafsson (1965), and of Archigetes sp. (order practical problems of taxonomy at the family, Caryophyllidea) by Motomura (1929). Where genus and species levels; another is to learn if comparison between species in the same genus such studies can help in appraising the rela- was made, especially of Hymenolepis by Jones tionship of these unusual tapeworms to the (1945) and Diphyllobothrium by Wikgren and more common strobilate groups, from which Gustafsson (1965), the chromosome numbers they differ greatly in morphology and general and/or karyotypes showed little variation and biology. therefore could be used little as an aid in the Hunterella nodulosa Mackiewicz and Mc- taxonomic analyses of the polyzoic cestodes Crae, 1962 (Cestoidea: Caryophyllidea) of the studied. family Caryophyllaeidae was chosen for de- From the single investigation of Motomura tailed cytological studies for two reasons. First, (1929), who was primarily interested in the in a preliminary survey of 17 species of caryo- embryology rather than the cytology of Archi- phyllids studied by the squashing technique, squashes of H. nodulosa had large numbers of 1 On academic leave from State University of New York at Albany, New York. good metaphase plates with relatively large, Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 36, NUMBER 1, JANUARY 1969 127 well stained chromosomes in the testes and acetic acid. A large proportion of the testes vitellaria. Second, H. nodulosa is easily identi- and vitelline follicles were teased out and, fied, widely distributed in North America, and along with the remaining tissues, dissected relatively easy to obtain, occurring as a com- into small fragments; only large pieces of the mon intestinal parasite of the white sucker, integument were discarded. A 22 mm square Catostomus commersoni (Lacepede). It is coverslip (No. 1 or 0) was applied and, after among the best known of North American excess acetic acid had evaporated, the prepara- Caryophyllidea with recent studies on syste- tion was squashed by placing the slide within matics, morphology, and distribution (Mack- one fold of paper towel and applying with the iewicz, I960; McCrae, I960; Mackiewicz and thumb the full weight of the body. The pres- McCrae, 1962) periodicity (Calentine and ence of Newton rings under the coverslip in- Fredrickson, 1965) and larval development dicated a satisfactory squash. The preparation (Calentine, 1965). was made permanent by using the dry ice method of Conger and Fairchild (1953), de- Materials and Methods hydrating in 95% or 100% ethyl alcohol for 1 Parasites were obtained on 27 November min and mounting with diaphane. 1967 from the white sucker, Catostomus com- Dividing cells were most common in the mersoni (Lacepede) (Cypriniformes; Cato- testes but excellent mitotic figures also oc- stomidae) from Jacks Creek, a tributary of the curred in the vitelline follicles, particularly in Smith River (Roanoke drainage), Patrick Co., cestodes in which spermatogenesis was just be- Virginia and from the same host from the Little ginning. Mitoses were rare in the parenchymal River, Walland, Blount Co., Tennessee on 29 cells. March 1968. Living cestodes were dropped Eighteen squash preparations representing into a vial of Carnoy's fixative (6:1:1, ab- a large number of testes and vitelline follicles solute ethyl alcohol, chloroform, glacial acetic from eight cestodes were studied. All illustra- acid) that was placed in ice water and later tions and most of the observations were made stored in a refrigerator until squashes were on squashes from immature worms collected in prepared in May, 1968. No differences in Virginia; the Tennessee collections consisted staining quality were detected in material of mostly gravid worms and were used to sup- stored for different periods. Carnoy fixation plement and verify the other observations. gave good spreads and preservation of the Camera lucida drawings of 16 metaphase plates chromosomes, unlike the results of Wikgren studied under oil immersion (900 and 1,800 (1964) who worked with plerocercoids of D. powers) were prepared at an enlargement of latum. 2,600 X. From 11 of these drawings of meta- Squashes were prepared using the leuco- phase plates (three from testes and eight from basic fuchsin (Feulgen) stain prepared ac- vitelline cells of two worms) an idiogram was cording to Darlington and La Cour (1960). constructed by measuring the chromosomes to Cestodes were removed from the vial of fixa- the nearest 0.5 mm and expressing each chro- tive, hydrated with successive changes (2 hrs mosome or arm as a percent of the total chro- each) of 95%, 50% (ethyl alcohol), and tap mosome length in the complement. In this way water. The whole worm was hydrolyzed in 1 there was an adjustment for the differences in N HC1 at 55 C to 60 C for 9-11 min (10 min chromosomes due to squashing or other factors. was later found to be a good average time) In those cases where either the centromere po- and removed to the Feulgen stain for one hr. sition could not be determined or the chromo- Usually the testes and ovary began to show a some was folded, no measurements were at- staining reaction within the first minute. When tempted; the sample size for each chromosome these organs became deeply stained, (usually measured therefore varied. The position of the within one-half hour), the whole worm was centromere of the small rods could not be ac- placed in 45% acetic acicl for 5-10 min in curately determined but from indirect evidence, order to soften and swell the tissues. A small as discussed under "observations," these chro- part of the body, approximately one-half mm mosomes appeared to be acrocentric and are so long, containing testes and vitellaria was placed indicated on the idiogram. Karyotype analysis on a slide and teased apart in a drop of 45% was based on the above eleven plates plus five Copyright © 2011, The Helminthological Society of Washington 128 PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY Figures 1-5. Chromosomes of Hunterella nodulosa. Feulgen stained squashes. 1, 2. Metaphase plates from vitelline cells. 3. Diakinesis. Note that more than one chiasma is readily visible on most bivalents. 4. Metaphase I, showing seven bivalents. 5. Metaphase II, Anaphase II, both dyads and monads are visible. others, only parts of which were suitable for ("J's") (Figs. 6, 7). At mitotic metaphase in purposes of an idiogram, that included two the spermatogonia and vitelline cells the long- from testes and three from vitelline cells of est chromosomes are 6—8 ^, the shortest 2.5-3 three worms (two from Virginia, one from ^ long. As shown by the idiogram (Fig. 6) Tennessee). there are four large "V's," two small "V's," two The photographs were made with a Zeiss large "J's," and three pairs of small rods. The Photomicroscope using phase contrast optics, largest chromosome represents approximately including an apochromatic oil immersion lens 12% of the total chromosome complement and a green filter. (TCL). No satellites or fragments were ob- served. The location of the centromere of the Observations mitotic metaphase chromosomes could be de- Hunterella nodulosa has 14 chromosomes, termined for all but the six rods, but from diploid (Figs. 1, 2, 7, 8). These include three observations of metaphase I of spermatogenesis pairs of metacentrics, ("V's", two long and one it appears that the centromeres of these rods short pair), three pairs of short acrocentrics are terminal or nearly so. ("rods"), and one pair of long submetacentrics Meiosis was studied only in spermatogenesis Copyright © 2011, The Helminthological Society of Washington OF WASHINGTON, VOLUME 36, NUMBER 1, JANUARY 1969 2N = 14 HUuu <f Figures 6-8.
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