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Histology of the Alimentary C*Nal of the Millipede

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Histology of the Alimentary C*Nal of the Millipede The histology of the alimentary canal of the millipede Orthoporus punctilliger Chamberlin Item Type text; Thesis-Reproduction (electronic) Authors Wilson, Barbara Eva Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 03/10/2021 03:00:29 Link to Item http://hdl.handle.net/10150/553155 HISTOLOGY OF THE ALIMENTARY C*NAL OF THE MILLIPEDE ORTHOPORUS PUNCTILLIGER CHAMBERLIN By Barbara Eva Wilson Submitted in partial fulfillment of the requirements for the degree of Master of Science in the College of Education of the University of Arizona 1932 ■ u "T:. v T '! ■ IF? 7 9/ /?32s 4 $ THE HISTOLOGY OF THE ALIMENTARY CANAL OF THE MILLIPEDE ORTHOPORUS PUNCTILLIGER CHAMBERLIN The study of the histology of the al­ imentary canal of Orthooorus punctilllger Chamberlin has been undertaken in an effort to add to the scanty litera­ ture on the anatomy of the diplopods, and to present some definite knowledge concerning an animal common to this vicinity. Acknowledgements are due to Dr, Hor­ ace Gunthorp for his supervision of the work, for his help­ ful advice and comments, and for the use of his literature on the Diplopods; and to Mr. Clair Hannum for his assistance in German translations, for his advice relative to histo­ logical technique, and for his help with the plates. Identification of Specimen This diplopod has been identified on the basis of Chamberlin’s description published in 1923. His specimens were collected in 1921, at Sonora, Mexico. Finding representatives of the species in Tucson is a new record in this State. Review of Literature The diplopod literature dealing with systematic classification is abundant, while that cover- 85800 2 ing anatomy and histological structure is very limited. The first work along the latter lines was that of Pla­ teau (1876), who studied the digestive systems of several chilopods and diplopods without sectioning methods, and therefore gave only very general pictures. In 1886, Keathcote presented the embryology of Julus terrestris. Little has been done in this field since, and his work stands as the foundation upon which most of the diplopod embryology is based. Relative to the alimentary tract, he found that the stomodaeum and the proctodaeum were formed by invaginations of the ectoderm, the midgut was formed from cells that remain within the yolk after the formation of the blastoderm, the malpighian tubules originated from the proctodaeum, and the salivary glands were mesodermal in origin. Krug (1907) concerned himself with the anatomy and histology of three species of diplopods, Blanlulus ^attulatus. Julus foetldus. and J. sabulosus, all European forms. His findings were confirmed or corrected by Ver- hoeff (1915) and Random (1924). Verhoeff published a mono­ graph on the diplopods of Germany in which he gave a com­ plete histology of their alimentary tracts along with a discussion of the mouth parts and head glands. He studied a great many forms, especially the Julidae, and often corrected Krug, but apparently he was not always right in his corrections. His histological studies form only a small portion of his entire work. Random made an intensive 3 study of the morphology and physiology of the alimentary . tracts of theJulidaeof which he studied nine different spedieei including these that Krug smirked on. In the 'maim 1 8e cleared up the points of difference between Krug ami ' Verhoeff,who were both found to be incorrect in certain ; details* fie w e t complete work bn head appendages is that of Snodgrass (1928),who worked out the morphology and evolution of the insect head* Hefner (1929) * by studying a single form. Paraiulua impressua, largely confirmed Ren­ dow *s findings* but added in addition a description of the head glands* In all these works there are minor paints of difference, but it must be b o m e in mind that these men are" working largely on different species and some variation ; should" be ■ expo©ted «■ -/ •'' •' ' ;■-1 : ; :: • Material and Methods -in’ Five specimens of Orthonorus punctilliger were used in this study, all of which were collected in the vicinity of Tucson* Arisona, where they are common during certain seasons of the year, fhey are most often seen early in the morning or late at night in the summer or spring months after it has rained. The latter fact accounts for the current name of "rainworm" given to the animal by the natives of the state! In cold weather they are very seldom observed, and probably live under .ground* - ' All animals obtained were mature and well- protected by a heavy exoskeleton, thus making it impossible 4 to sootldh the whole body, as Hefner did with Para.Itilua la* prosaus. After the animals had been collected, some were killed v/ith ether or cyanide and disseoted immediately, while ethers were killed in Bonin's fixative and disseoted at a later date • The first method proved most satisfacte^^ as the gut could bo removed In a much better condition. Oh the whole dissection was a difficult and tedious task due to the hard exoskeleton of chitin which covers the animals, and to the intricate manner in which the cnfrali are overlapped. The exoskeie^On lies very close to the alimentary tract and in removing it care must be taken to prevent serious injury to the gut. This is especially true when the gut Is dlstended with food. The animals invariably die in a spirally coiled condition, but this caused ho trouble. ; Gross dissection was accomplished by cutting a long V-shaped groove out of the paraffin in a dis­ secting pan, and placing the specimen in the groove in such a manner as to leave the dorsal surface free from’ -and above the level of the paraffin. The sides of the paraffin were then melted down over the legs of the animal, and if necessary, other melted wax was poured into the groove and allowed to harden. In this way the animal was firmly held, in position by the paraffin which covered its appendages and its ventral and ventre-lateral surfaces. The wak was thoroughly hardened by immersion in water, and the exoskeleton cut along the lateral margins as low down as possible with a knife or a small pair 5 of scissors• As soon as both sides of;the a nmiM had been severed, the entire dorsal half uas lifted off. Due to the intricate overlapping of the annuli, it was discovered that if a third and dorsal out were mad®, the rings of the exo- skeleton eould be more easily removed in two pieces than in one. All dissection was done under a normal saline solu­ tion. The more difficult cuts were performed with the aid of a dissecting microscope. Upon the removal of the alimen­ tary canal, it was cut into pieces about half an inch long, and thoroughly washed. The latter is necessary because small pieces of sand often get lodged betimen the folds in the gut. Qrthoporus puhctilliger is a vegetable feeder, but it lives underground and consequently gets grit into the digestive tract. The tissues were fixed In Bonin's fluid for twelve hours, a M toen transferred to 70^ alcohol to which a little lithium carbonate had been added. When all the picric acid had been removed, the tissues were de- hydrated and cleared by running them through mixtures of butyl and ethyl alcohol in the following proportions: 1. 40 parts 95f0 ethyl §0 parts water — 50% 10 parts butyl 2. 50 parts 95% ethyl 25 parts water - - - - - 70% 20 parts butyl 3. 60 parts 95% ethyl 10 parts water - - - - 9U% 30 parts butyl 4. 60 parts 95% ethyl 7 ; ' 40 parts butyl » » - 1CXK V - 5. SO parts of 95^ ethyl ' . m , • 70 parts butyl - - - 1QO& , . , 100 parts butyl - - - 100^ ; At first the, tissues were left in the grades of alcohol for a full hour. #aoh*, but it was found that the hindgut and .. for®gut» because ®f their structure, tended to harden and made sectioning Impossible, so the time was. decreased to , thirty minutes each for the first four changes and forty- five for the last two, and better results were obtained# While the tissues, were in the absolute butyl alcohol,, a vial was filled two-thirds full of melted paraffin, and allowed, to stand for a short time. The tissues were placed,on the hardened paraffin, covered, with butyl alcohol and left in the oven until they sank to the bottom of the vial# They then were transferred^into the first of four changes of melted paraffin in which they remained for thirty minutes each. Care was taken to see that all butyl alcohol was displaced from the tissues* The material was embedded . in hard paraffin (54° to 56® C), mounted in the usual way for paraffin blocks, and sectioned# , , Hcmy difficulties,were experienced, in sectioning due to the bard ^ chitin in the hindgut and fore- , gut# In an attempt to overcome this, the pieces, of the alimentary oanal were placed in 10^ nitric acid,, but the results were not wholly satisfactory.because the tissues 7 wore found to become very brittle and- hard as ‘they were : ' run through the aloohola. A aeccmd a e t h M eonsioted in rihe- tng' the'"tissues;In'TOjC aloohol «td then placing thea in dia- ‘ phenol until thoroughly bleached and softened. After they had been hardened in 70^ alcohol, they were transferred through tetralln into* paraffin. This method helped to a certain ex­ tent.
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