Ehrlich's Acid Research Articles

The Life Cycle of Plagiorchis vespertilionis parorchis, n. ssp., (Trematoda: Plagiorchiidae), and Observations on the Effects of Light on the Emergence of the Cercaria

Many Lymnaea stagnalis L. collected from Cascade Lake, Orcas Island, Washington, were found to shed large numbers of a species of xiphidiocercaria containing refractile granules. They resembled the cercariae of Plagiorchis as described by McMullen, 1937, and subsequent experiments proved that they were the larval stages of what appears to be an undescribed subspecies of Plagiorchis vespertilionis (Miiller, 1784) Braun, 1900. Encystment was induced experimentally in trichopterous larvae, dragonfly nymphs, ephemerid larvae, and Culex mosquito larvae. All were collected in the vicinity of Portland, Oregon, where Lymnaea stagnalis is not known to occur, and a series of such insects were dissected to demonstrate that all were free of trematode cysts of this type. Adult flukes were obtained by feeding infected insects to white mice, and proven to be a common species found in small Myotis bats on Orcas Island. This work was supported by Contract Nonr 1805(00), NR 130-342, between the Office of Naval Research, Department of the Navy and Portland State College. The writer also wishes to acknowledge the assistance of Donald J. Moore for his help with certain aspects of the investigation. Further he is under special obligation to Dr. Franklin Sogandares-Bernal for furnishing a series of drawings of the position of organs in Plagiorchis vespertilionis from Korean bats. Dr. Murray L. Johnson of Tacoma, Washington, kindly identified the bats, specimens of which numbered 6922 to 6937 are now in the Puget Sound Museum of Natural History. METHODS Snail hosts transported in glass jars aerated with oxygen generators survived the 300-mile trip from Orcas Island to Portland in excellent condition. In the laboratory they were separated into shallow white-enameled pans covered with glass. The snails survived and could be reared from eggs in such pans if the bottoms were provided with fine gravel and water was maintained at a level not much higher than the diameter of the snails. Food was provided by maple leaves first placed for a minute in boiling water to soften them. Tap water which had been allowed to stand in large jars for at least 24 hours was quite satisfactory as a substitute for pond water. To check snails for infections, they were isolated overnight in shell vials 2 by 9 cm in size. Cercariae were studied alive with and without neutral red stain. Those used for measurements first were lightly stained with neutral red and then killed in hot 10 percent formalin. Adult trematodes were fixed in Gilson's fluid after slight compression under a cover glass, and then stained with borax carmine and fast green or with Ehrlich's acid hematoxylin. For the experiment on the effect of reversal of light and dark periods, light-tight cabinets were employed. These were furnished with fluorescent lights to minimize temperature changes and constructed for adequate ventilation. The lights were controlled by automatic time switches. Measurements are in millimeters unless otherwise indicated. Figures 1 to 15 are of P. vespertilionis and were drawn with the aid of the camera lucida.

Histology and Cytochemistry of Skin Gland Areas in the Caribou, Rangifer

The microscopic structure and glandular cytochemistry of three specialized cutaneous areas in the caribou form the subject of this report. These are the tarsal gland area, the interdigital gland areas and the preorbital pocket. Their location and gross structure may be seen in Plate I. These structures have been shown to vary in their presence and structure among artiodactyls, and have been most recently reviewed by Schaffer (1940). There is still much, however, that is unknown concerning them, particularly regarding seasonal and sexual variation and microscopic structure and chemistry of the glands. It is hoped that this report will help fill these deficiencies for the caribou, and provide significant information for behavioristic and phylogenic studies as well as for comparative glandular cytology and cytochemistry. I am very grateful to Ralph S. Palmer for obtaining and providing the skin samples upon which this study is based. Financial support for publication was provided by funds derived from income on the endowment of the Horace H. Rackham School of Graduate Studies, University of Michigan. This study is based upon skin samples from an adult male and an adult female, both shot at Anaktuvik Pass, Brooks Range, Alaska on April 3, 1950. The skin samples were fixed in a formalin-alcohol mixture and were later stored in 70 per cent alcohol. Blocks of tissue, the locations of which are indicated in Plate I, were cut from the samples, washed, dehydrated through a series of ethanol solutions to benzene, and embedded in paraffin. Serial sections 10 microns in thickness were treated by the following techniques. (1) Ehrlich's acid hematoxylin and eosin Y (Bensley and Bensley, 1938). (2) Taenzer-Unna acid orcein method for elastic fibers (Lillie, 1948. The orcein, dissolved in 70 per cent ethanol was used at room temperature. Azure II (Nat. Aniline …

The use of polyvinyl alcohol and its derivatives as microscopical mounting media. Part II. Hydrocarbon-miscible mounting media.

SYNOPSISA mounting medium with a refractive index of 1·474 had been developed from a 23 p.c. dispersion of polyvinyl acetate in “cellosolve” plasticized with 13 p.c. of diamyl phthalate. When heated the medium hardened in 2–3 days to a stable transparent film which did not cause stain‐fading in test mounts stained with either Ehrlich's acid alum haematoxylineosin B or with Mallory's acid fuchsin‐orange G‐aniline blue and exposed for 6 weeks to heat, artificial, or natural lighting.

Sanguinicola huronis n. sp. (Trematoda: Sanguinicolidae) from the Blood System of the Largemouth and Smallmouth Basses

During a parasite survey of northwest Wisconsin fishes, a species of blood fluke of the genus Sanguinicola Plehn, 1905, was reported by the author (1947) from the mesenteric blood vessels of the largemouth bass, Huro salmoides (Lac.) and the northern smallmouth bass, Micropterus d. dolomieu Lac. In the 1947 paper, without naming or describing the fluke, it was declared a new species because it differed morphologically from the single known North American species, Sanguinicola occidentalis Van Cleave and Mueller, 1932, from the heart of the walleye, Stizostedion vitreum. The genus Sanguinicola was established by Plehn (1905) to receive S. armata and S. inermzis from European cyprinids; she called them endoparasitic Turbellaria. Later (1908), she reviewed her earlier conclusions and reclassified the two species as Cestodaria. Odhner (1911) correctly placed these forms in the Trematoda and discussed his and Looss' observations on the life cycle of S. inermis. The life cycle of this species was further clarified by Scheuring (1922). Woodland (1923) described a species of Sanguinicola from a Nile silurid, but did not name it. Odhner (1924) redescribed this species and named it S. chalmersi. S. intermedia was described by Ejsmont (1926) from European cyprinids. He also presented information on S. armata, S. inermis and S. chahnersi as well as additional life cycle data. Van Cleave and Mueller (1932) described S. occidentalis from a percid fish, the walleye (Stizostedion vitreum), in Oneida Lake, New York. More information on the biology and ecology of this species was presented by Van Cleave and Mueller (1934). Fischthal (1947) found S. occidentalis fairly widely distributed in the walleyes of northwest Wisconsin, and also reported this species from an additional percid host, the yellow perch (Perca flavescens). The present species was first observed April 15, 1944, when six adults were recovered from one of 11 Huro salmoides collected from Bear Lake, Barron County. On September 20, 1944, seven of 12 Huro salmoides from Lost Land Lake, Sawyer County. were found parasitized, two with one worm each and five with two each. Also on this date, seven worms were recovered from one of two Micropterus d. dolomieu from Lost Land Lake. On October 4, 1945, a fingerling Huro salmoides from Spring Creek, Washburn County, was infected with one worm. The author recalls finding in 1940 a single, typically triangular Sanguinicola egg in the ureter of Huro salmoides from the Huron River, Washtenaw County, Michigan. In all probability it belonged to the species herein described. The name Sanguinicola huronis is now proposed for this species. Its description is based on whole mounts of specimens fixed in AFA, stained in Ehrlich's acid haematoxylin, and mounted in balsam. The interpretation of the morphology presented many difficulties as encountered by other workers on the genus Sanguinicola.

Triganodistomum hypentelii n. sp. (Trematoda: Lissorchiidae) from the Hog Sucker, Hypentelium nigricans (Le Sueur)

Examination of the small intestine of a hog sucker, Hypentelium nigricans (LeSueur)2 collected on April 3, 1941, from the Saline River near Milan, Monroe County, Michigan, revealed three specimens of a new species of trematode belonging to the genus Triganodistomum Simer, 1929. In the fall of 1941, two of nine hog suckers taken from the Saline River near Saline, Washtenaw County, Michigan, contained three and four adult trematodes respectively. The following description is based upon the study of living specimens, and of whole mounts and transverse sections of specimens fixed in hot Bouin's fluid, stained in Mayer's paracarmine (whole mounts) or Ehrlich's acid haematoxylin (sections), and mounted in clarite X (Nevillite no. 1).

A New Astomatous Ciliate from the Newt, Eurycea bislineata (Green)

In the course of examination of the intestinal contents of 21 specimens of Eurycea bislineata (Green) collected at Oxford, Mass., during the summer of 1939 a single newt was found of which the small intestine was heavily infested with an astomatous ciliate. Observations and measurements were made on the living organisms which were later killed with hot saturated aqueous mercuric chloride solution containing 2% of acetic acid. These specimens were ultimately stained, some with acid carmine, others with Ehrlich's acid haematoxylin with or without a counterstain of eosin.

Hymenolepis scalopi n. sp.

The worms were studied both alive and preserved. They were fixed either in Gilson's or Bouin's fluid, stained in borax carmine, cleared in toluol, and mounted in balsam. Some of the pieces were later removed from the balsam, sectioned, and stained in Ehrlich's acid haematoxylin. All of the measurements recorded below, except the length and width of the entire strobila, were taken from preserved specimens.

THE INVARIABLE BLOOD-STAIN

For years I have used in my laboratory a method for the staining of blood-films which has proved so satisfactory from all points of view as to have earned the term invariable. The idea is not original with me, but is based on two well-known principles, combined, modified and finally perfected. First of all, single films are best fixed by burning in absolute alcohol; a proper mixing of eosin with Ehrlich's acid hematoxylin gives an excellent stain, one which proved its worth before the panoptic formulas became so popular. This method is especially suited to the needs of the general practitioner, the man who desires results and who cannot become proficient in technic. Certain men, who do considerable blood work, are now using the invariable almost routinely. Several of these have prevailed on me to publish the formula and technic; and this will perhaps justify the addition of another