Rana Palustris Research Articles

A Range Extension of the Pickerel Frog, Rana palustris palustris, in Oklahoma with Indications of an Extension of the Austroriparian Life Province

A Range Extension of the Pickerel Frog, Rana palustris palustris, in Oklahoma with Indications of an Extension of the Austroriparian Life Province

High Incidence of Two Parasitic Infestations and Two Morphological Abnormalities in a Population of the Frog, Rana palustris Le Conte

Adult frogs, Rana palustris, trapped at a small pond during three breeding seasons, were infested by larvae of a mite, Hannemania dunni, with annual frequencies of 60%, 90%, and 83%, an infection rate greatly exceeding that encountered in other anurans inhabiting the same pond. Ninety normal juveniles were trapped in 1961. In 1962, 990 juveniles were trapped. In a selected sample of 45 of the latter, 39 had heavy infestations of spiruroid nematode larvae, 12 had incompletely erupted forelimbs, and 4 had unflexed hearts. No such conditions were seen in juveniles of other species nor were. juveniles of Rana palustris so affected in 1963. Possible causal interrelationships of these conditions are suggested.

Nucleocytoplasmic hybrids between Rana pipiens and Rana palustris: I. Analysis of the developmental properties of the nuclei by means of nuclear transplantation

Although diploid hybrids between Rana pipiens and Rana palustris are normal, the androgenetic haploid hybrids develop much more poorly than do ordinary haploids of either species. This suggested the existence of a nucleocytoplasmic incompatibility which is expressed when a nucleus containing chromosomes from only one of these species is present in the cytoplasm of the other. Since haploids, in any case, are not viable, a valid test required the study of diploids. Such a study, as reported in this paper, demonstrates that this nucleocytoplasmic incompatibility does in fact exist. When diploid pipiens blastula nuclei were transplanted into enucleated palustris eggs, the resulting nucleocytoplasmic hybrids displayed a specific pattern of deficiencies leading to their developmental arrest in postneurula stages. Whether or not this incompatibility causes stable changes in the developmental properties of the nuclei was then tested by transferring pipiens nuclei after division for one, two, and three blastula generations in palustris cytoplasm back into enucleated pipiens eggs. The large majority (84%) of the back-transfer embryos developed normally, thus demonstrating that the nuclei remained unchanged in their ability to function normally in their own cytoplasm following as many as 36 divisions in the foreign egg cytoplasm. The possibility that such changes might take place later in the development of the nucleocytoplasmic hybrids remains to be worked out. Cytological analyses of the nucleocytoplasmic hybrids and the back-transfer embryos revealed that both types of embryos possessed normal chromosome complements. This result is particularly significant with regard to the nucleocytoplasmic hybrids since it shows that their deficient development is not caused by gross chromosomal abnormalities. In addition, work described elsewhere shows that the abnormal behavior of these embryos can be completely corrected by the addition of a haploid set of palustris chromosomes. Thus, this hybrid combination offers a unique opportunity for analyzing genetic and biochemical events involved in the formation of a predictable set of developmental deficiencies in embryos whose nuclei have normal chromosome complements and are known, under defined conditions, to be genetically competent.

The Occurrence of the Pickerel Frog, Three Salamanders and Two Snakes in Mississippi Caves

The Occurrence of the Pickerel Frog, Three Salamanders and Two Snakes in Mississippi Caves

Abnormal Combinations of Nuclear and Cytoplasmic Systems in Frogs and Toads

This chapter illustrates the development of genetic differences that lead to hybrid inviability. It is pointed out that the species level of divergence can be reached with no hybrid inviability at any stage of the divergence. For example, rana pipiens and rana palustris are full species, and no herpetologist regards them as anything else. Their hybrids are entirely normal, so there is no reason to suppose that hybrid inviability was ever a concomitant of their divergence. It is probable that there is no necessary relation, in allopatric populations, between hybrid inviability and evolutionary divergence at the lowest levels. When two populations are evolving in different regions, the thing of primary evolutionary importance is the adaptation of each to its own environment. If the two happen to adapt in different ways, they are said to diverge. The genetic basis of this adaptation may or may not be of a sort that leads to hybrid inviability if the two are crossed. This divergence need not involve the development of different external characters, and if it does not, the chances of detecting it are reduced. There is no reason that adaptation should involve the development of differences that can be used conveniently by taxonomists.

STUDIES IN THE DEVELOPMENT OF FROG HYBRIDS. IV. COMPETENCE OF GASTRULA ECTODERM IN ANDROGENETIC HYBRIDS

1. In an effort to dissociate the factors responsible for the failure of development in Rana pipiens ♀ x Rana sylvatica ♂ hybrids, a study has been made of the development of haploid embryos composed of pipiens cytoplasm and sylvatica chromosomes. These androgenetic hybrids develop only to the late blastula stage.2. The competence of the presumptive ectoderm of these haploid hybrids was tested by transplantation to neurulae of Rana palustris. The results indicated a total lack of competence.3. Aside from the main problem with which the paper is concerned, it was found that the various classes of embryos used could be arranged in order of decreasing competence of the presumptive ectoderm. The sequence was as follows: diploid pipiens, haploid pipiens, pipiens ♀ x sylvatica ♂ diploid hybrids and (pipiens ♀) x sylvatica ♂ haploid hybrids.

Hybridization Between Rana Palustris and Different Geographical Forms of Rana Pipiens

Hybridization Between Rana Palustris and Different Geographical Forms of Rana Pipiens

A Mutant Red-Phase Wood Salamander (Plethodon cinereus) from New Hampshire

This is a report on several series of amphibians and reptiles, received from New Hampshire during the spring of 1945. The most interesting of these specimens were eight wood salamanders from Andover, Merrimack County: seven of the common red-backed phase described by Bishop (1943, Handbook of Salamanders, p. 234), none of the lead-backed or dark phase, and one of a new all red phase not mentioned by Bishop. The red mutant was typical of Plethodon cinereus in every way, except for lack of dark pigment on the sides and below. It was uniform light red from the lower flanks upward and light below. The collector of this specimen was Miss Katherine Gulick, who also sent from the Andover area: Desmognathus fuscus fuscus, dusky salamander; Eurycea bislineata bislineata, two-lined salamander; Triturus viridescens viridescens, eastern newt; Hyla crucifer, spring peeper; Rana palustris, pickerel frog; and Chrysemys picta picta, eastern painted terrapin. Additional New Hampshire records from 1945 collecting: young Hyla crucifer taken at Raymond, Rockingham County, by Robert A. Hellman; water stage Triturus viridescens viridescens found at Meredith, Belknap County, by Peter Lunt and the red eft land stage at Webster Lake in Merrimack County by Irene Wilson; and the milk snake Lampropeltis triangulum triangulum secured at Andover, Merrimack County, by Winslow Trueblood.

Developmental Rate of Hybrids between Rana pipiens and Rana sphenocephala.

This forms the second report on developmental rate in hybrid frogs. The purpose of these experiments, in hybridizing species that differ in rate of development, is to determine the earliest effect of the sperm on this particular character. Previously such a study has been made on hybrids between Rana pipiens and Rana palustris.1 Both hybrids, pipiens ♀ × palustris ♂ and palustris ♀ × pipiens ♂ developed at the maternal rate during early stages. The first unmistakable deviation from the maternal rate appeared in the neural plate stage in both hybrids. Additional evidence on these points is derived from the following crosses.The material used in these experiments is from the following sources: Rana pipiens Schreber from dealers in the Lake Champlain area of Vermont and Rana sphenocephala (Cope) from dealers in southern Florida. Although listed as separate species by many taxonomists the two are closely related and are thought best included under one name by Kellogg.2 Regardless of their true taxonomic statu...

The phenomena of anuran metamorphosis. III. The development of the thyroid gland

AbstractThis paper presents a study of the growth and histological development of the thyroid in three species of frogs, the small Pseudacris triseriata, the large Rana catesbeiana and the intermediate Rana palustris. It was found that the history of the gland in relation to metamorphosis is essentially the same in all. Previous to metamorphosis the gland is small, and the cells very poor in cytoplasm. At the beginning of metamorphosis (i.e., rapid hind leg growth) the thyroid shows a marked acceleration of growth and activity, which continues and reaches a peak at the time of the emergence of the front legs. Late in metamorphosis a regression occurs.The conclusion is drawn that there is a rising concentration of thyroid hormone during the course of metamorphosis. The author has previously shown that such a rising concentration will induce the normal pattern as to sequence and time spacing of metamorphic events.The beginning of metamorphosis is conceived as dependent upon the time of stimulation of the thyroid and the pattern of its subsequent continuance as dependent upon the pattern of response of this gland.

Experimental Formation of Accessory Organs in Mid Body Lateral-Line of Amphibians.

The ability of primary lateral-line organs in amphibians to form accessory ones by budding,, the time at which these bud organs are developed, and the pattern of organ groups gave a method for measuring the effects of the present experiments in amphibians. When the placodes of the mid-body lateral-line primordia of young and older tail-bud stages of Amblystoma punctatum (Harrison stages 23-24 and 28-29) are exchanged, the age difference between graft and host organ-forming tissue can be clearly followed through the young larval stages by observing the time at which accessory organs are laid down by the budding of primary ones. In A. punctatum the dorsal and ventral poles of primary organs give rise to accessory organs forming lineal groups of 2 or 3. In A. tigrinum by a similar process groups of from 4 to even 8 are formed, usually in irregular clusters, as in Rana palustris and Hyla crucifer. There is multiple budding of both primary and accessory organs. When the mid-body lateral-line primordia are exchanged in the tail-bud stages of punctatum and tigrinum the formation and grouping of organs from the graft follow only that of the donor species. When the lateral-line primordium of A. punctatum is placed on the body of A. tigrinum in the future pathway of the primordium of the host, the one of the graft will migrate and form organs only when the primordium of the host touches and fuses with similar material of the graft. The phenomenon is similar to that previously reported. At the appropriate times accessory organs are developed, which in size and group formation are identical with those of the donor species. The organs are innervated by the host lateral-line nerve and never were observed to degenerate up to late larval life.

Production and Metamorphosis of Chimeras in Anurans and Urodeles

The perfect union of the anterior and posterior halves of the embryos of 2 species of anurans and the metamorphosis of such a combination into a perfect adult was shown to be possible by Harrison 30 years ago. Up to the present day the opportunities which this operation makes possible have never been realized. The intention of the present report is to demonstrate that such combinations can be produced in large enough numbers to enable the experimental embryologist and geneticist to carry out a variety of new studies. The frog embryos employed were Rana palustris and Rana sylvatica. Those of the salamander used were Amblystoma tigrinum and Amblystoma punctatum. The stages at which the operation was done were in the early tail-bud period of development. The species conbinations in their respective classes are, at the tail-bud stages, about the same in size. Similar stages in development were selected and placed in slightly diluted Ringer's solution in an operating dish, the floor of which was covered by a thick film of white beeswax. The embryo was oriented under the microscope, dorsal side up, between fine forceps so that iridectomy scissors could, with one stroke, cut the embryo transversely at the level of the seventh to eighth somites. It is quite possible to cut at other levels as well in making successful combinations. The anterior half of one species and the posterior half of the other were then placed in a depression in the wax which was about the size of an intact embryo. The cut surfaces of the reciprocal halves were brought together in the concavity of the wax as soon after the operation as possible.

EFFECT OF RINGER'S SOLUTION ON TAIL-REGENERATION IN THE TADPOLE OF RANA PALUSTRIS

Tadpoles of Rana palustris, regenerating new tails, produce, in general, less new tissue when kept in an approximately isotonic frog Ringer solution than when allowed to remain in ordinary fresh water.The regeneration of the caudal membrane is frequently affected by isotonic solutions. Occasionally the bud of new tissue consists merely of the main axis without any membrane. In some other cases a much reduced or irregular membrane is formed.

Developmental Changes in Primary Lateral-Line Organs Studied in Living Larvae of Anurans and Urodeles

In amphibian embryos the migrating lateral-line primordia lay down a series of primary organs which can be observed in the living and vital-stained condition.1, 2 Since the mid body line extends down the tail in rather transparent tissue such organs are favorable for study in the living specimen when observed under high magnification in micro-aquaria. These data concerning the developmental changes in primary tail organs of normal larvae provide a basis for further observations consequent to the cutting of the lateral-line nerve. In Amblystoma punctatum larvae the large melanophores often obscure the view of many lateral-line organs and their nerves. This condition was easily avoided by feeding specimens an exclusive diet of enchytraea which, for some unknown reason, contracts the dark pigment cells. The animals become pale yellow in color. In Hyla crucifer tadpoles pigment presents no serious difficulties. In contrast to the normal pigmented larvae of Rana palustris it was found that hypophysectomized specimens gave as clear a picture as Hyla. In Amblystoma punctatum the tail organs begin abruptly at a level above the anus. In many larvae studied at the feeding stage (Harrison stage 46, 15-16 mm.) such a line on the tail is about 18 in number, extremes being 13 to 20. As in other lines the number on the right side is usually not equal to that on the left. From primary organs accessory ones are formed by budding from supporting cells.1, 2 Shortly after the feeding stage begins the terminal organ may give off a bud posteriorly and thus form a chain of 1 to 3 new organs. However, normal isolated animals often do not show an immediate response by budding of the terminal organ until later.

Lens Induction in Ectoderm Transplanted from Rana Palustris to Amblystoma Punctatum

Geinitz showed that, if the dorsal lip of the blastopore is transplanted from an anuran gastrula to the gastrocoel of a urodele, it not only differentiates into typical axial organs but also induces the formation of a neural tube and somites in the urodele tissue with which it is in contact. Lewis attempted to obtain lens induction by implanting an anuran optic vesicle beneath epidermis in the region between the eye and the ear of an Amblystoma embryo but he himself was not sure of the source of the lenses which developed after this operation. Since Geinitz's demonstration of xenoplastic induction, renewed attempts have been made to show that lens formation can be induced by xenoplastically transplanted optic vesicles. Cotronei and Spirito and Mangold have performed experiments bearing on this subject but have not obtained positive results in the species which they used. In the experiments here reported, Rana palustris and Amblystoma punctatum were used as donor and host respectively. The operation involved removing epidermis from over the left optic vesicle of the urodele host at stage 22-23 and substituting for it a piece of head epidermis from the anuran donor. The donor, which was at approximately the same stage of development as the host at the time of operation, was, however, considerably smaller than the host. Therefore, in order to cover the host's optic vesicle, it was necessary to transplant an extensive piece of the donor's head epidermis. Animals which had been operated upon were preserved at various stages during embryonal and early larval life, i. e., from 4 to 23 days after operation, and were subsequently examined histologically. Xenoplastic lens induction was found to have taken place in 11 cases.

Experiments showing the role of migrating neural crest (mesectoderm) in the formation of head skeleton and loose connective tissue in Rana palustris.

Experiments showing the role of migrating neural crest (mesectoderm) in the formation of head skeleton and loose connective tissue in Rana palustris.

On Specificity of Opalina in the Frog.

It is well known that the ciliate, Opalina, parasitic in the rectum of the leopard frog, Rana pipiens, and in the pickerel frog, R. palustris, occurs but rarely or not at all in the closely allied adult green frog, R. clamitans, and bull frog, R. catesbiana. Tadpoles of R. clamitans have been found by us to be almost universally parasitized. Experiments by the first author were conducted to determine if Opalina would live and reproduce when transferred directly from the rectum of R. pipiens to that of the two genera of green frogs. The frogs used in these experiments included R. pipiens, R. clamitans, and R. palustris, collected within 70 miles of New Brunswick, N. J., and R. catesbiana from New Orleans, La. The parasite employed in these transfers was O. obtrigonoidea. Removal of the rectal contents was accomplished with the aid of a fine glass pipette introduced through the cloaca of the host after previously relaxing the surrounding muscles by the action of M/8 magnesium chloride for 20 minutes. A pipette full of Ringers solution was first forced into the rectum before withdrawing the contents for examination or for introduction into a second host. The experiments were carried on from October to late May of the following year. The green frogs and bull frogs used in these experiments were examined in each instance prior to inoculation. One specimen of clamitans was found to harbor a heavy infection of Opalina, due, possibly, to contamination from close proximity to pipiens and their faeces, since the former frog does not normally harbor these parasites.

STUDIES ONEUGLENAMORPHA HEGNERIN. G., N. SP., A EUGLENOID FLAGELLATE FOUND IN TADPOLES

1. Euglenamorpha hegneri is the name proposed for a new Euglena-like flagellate found in the rectum of tadpoles and first reported by Hegner ('22).2. Most of the tadpoles examined were those of the bullfrog obtained from the pond in the Botanical Gardens of the University of Pennsylvania. Some tadpoles of Rana palustris collected by Dr. C. L. Parmenter and fed on vegetation from the same pond were also examined. About 60 per cent. of the 117 bullfrog tadpoles examined in 1922 and 16 per cent. of the 30 examined in 1923 harbored Euglenamorpha. About 60 per cent. of the 9 tadpoles of Rana palustris examined in 1922 and also of the 35 examined in 1923 were infested with this flagellate. Eight tadpoles of R. clamitans(?) and 8 of Hyla versicolor(?) examined at Woods Hole in August, 1923, all contained Euglenamorpha.3. There are two varieties: one, green, is the type, and the other, pellucida, is a colorless derivative. Typical individuals of the two varieties are so different that without intermediate stages they would be put into different species or into different genera.4. The green (type) variety has a cylindrical or cigar-shaped body, chlorophyll, stigma, paramylon granules, compact nucleus, three flagella with swellings on their roots near the stigma, surface striæ passing from the left over to the right. It may occasionally divide by mitosis within the host.5. Variety pellucida has an elongately conical body, little or no chlorophyll, no stigma, no swellings on the usually four or six flagella; has enlarged reservoir, nucleus expanded, and surface striæ usually passing from right over to the left or longitudinal. Amitosis as well as mitosis occurs frequently in the host.6. Transitions between the two varieties are found and it is believed that the green variety transforms into the colorless one. Division of the green variety in the host is rare, but appears to be accompanied by transformation which may also take place without division. Once established, the colorless variety may multiply rapidly by both mitosis and amitosis and consequently outnumber the green one.7. Attempts to culture the flagellates outside the host were more successful with hanging drops than with dishes. The most successful culture was a hanging drop of 0.6 per cent. NaCl in which the green variety multiplied for three weeks, then gradually declined, but some few were still alive after five months. In other cultures a few appeared to transform from the green to the colorless variety.8. The movements of E. hegneri, both swimming and metabolic, may be very rapid. In some cases the organisms appear to attach themselves to the wall of the rectum or other object by one flagellum to vibrate the others near the body, thus possibly serving both a respiratory and a nutritive function. They are positively phototropic.9. Outside the host (in culture) the green variety may assume a resting state approximating encystment in which it may divide and by means of which, presumably, it reaches a new host. The colorless variety is not known to assume the resting state and appears to be too unstable to be a permanent form.10. One is tempted to regard the entire situation as revealing an uncompleted series of stages in the evolution of a colorless parasite from a free-living, green euglenoid flagellate, but the evidence is not entirely conclusive.

Cytamoeba bacterifera in the Red Blood Cells of the Frog

This note is intended to call to the attention of parasitologists and bacteriologists the presence in the red blood cells of frogs in this country of a most interesting organism to which the name Cytamoeba bacterifera was given by Labbe in 1894. Cytamoeba bacterifera was found in the red cells in the peripheral blood of two specimens of Rana clamitans and five specimens of R. catesbiana collected at Cold Spring Harbor, Long Island, N. Y., in the summer of 1919. It was absent from the other frogs and toads examined; namely, Rana palustris, R. sylvatica, R. pipiens, Hyla versicolor, Bufo americanus and Scaphiopus holbrookii. In every case the host was also parasitized with trypanosomes, hemogregarines, and several other forms that represent either undescribed species or stages in the life cycles of parasites known only as adults. Freshly drawn blood was studied as well as material fixed and stained by Wright's method and by the Schaudinn-iron-haematoxylin method. The frogs were kept alive in the laboratory for a considerable period and their blood examined at intervals of a few days; several of them were kept in this way for three months.

Comparative study of ethanol, caffeine and nicotine on the development of frogs' larvae

The effects of ethanol, caffeine (alkaloid) and nicotine (alkaloid) solutions were studied on the growth and development of the larvæ of two species of frogs, viz., Rana sylvatica and Rana palustris. The study of the larvæ was begun immediately after hatching from the eggs and continued on tadpoles of older ages. Tadpoles of the same species and ages were placed in solutions of the above drugs of various concentrations and the effect of the poisons was noted. It was found that the toxicity of the drugs varied with the age of the tadpoles and with the nature of the drug. Very young tadpoles succumbed much more quickly to all the poisons than the older ones when placed in solutions of the drugs of the same concentration. Tadpoles 1 and 2 days old quickly died in all the solutions unless the drugs were present in a very dilute concentration. Older tadpoles continued to live in solutions of all the drugs longer than was at first expected. The most toxic of the 3 drugs in point of dosage was nicotine. Next in toxicity came caffeine and the weakest of the 3 drugs studied was ethanol. It was surprising to find that while caffeine even in great dilution proved deleterious to the growth and development of the tadpoles, ethanol affected the animals to a much lesser degree. Thus it was found that while tadpoles of the age of 8 days when placed in a solution of nicotine, 1-50,000 succumbed on the 23d day, other tadpoles of the same age placed in caffeine solution, I-10,000 died on the 12th day, while still other tadpoles of the same age placed in a solution of ethanol, 1-100, by volume, lived as long as 40 days.