(Katlilaniidae), and various tylenclis, hoplolainis and cricoiie- 

 matids and Dori/laimus stagnalis. 



Stored Protein. Such substances occur as non-birefiingent 

 colorless globules similar to fat globules in transmitted light 

 and dark field illumination. As described by Chitwood and 

 Jacobs (1937), they are insoluble in water, alcohol, xylol, ether, 

 and Vi-saturated ammonium sulphate; dissolve in 10% acetic 

 acid and in 5% KOH ; are not affected by saliva ; are pale 

 yellow in Flemming's fixative; give positive xanthoproteic and 

 ninhydrin reactions; and are digested by artificial gastric 

 juice. The globules stain with gentian violet or haematoxjdin; 

 they also stain blue with Nile blue sulphate and orange with 

 Scharlach R. These reactions apparently place them as com- 

 plex proteins of a con,iugated nature. The majority of glo- 

 bules of Agamermis are composed of this type of substance 

 and not fats as is commonly supposed. Similar globules have 

 been identified by the writers (1938) in the intestine of Dity- 

 len.e]nis dipsaci (Fig. lOOC). It is quite possible that the 

 colorless, insoluble (in 'Icoholxylol), basophilic globules, pres- 

 ent in Bhabditis, Ponafirolaimus, Aphelenchoides and Plectitx 

 etc., are of the same nature. The fact that such globules stain 

 with Nile blue sulphate in the same manner as fatty acids 

 indicates that staining technics are not necessarily indicative 

 of fat. Proteins may stain as do fats but they may be dis- 

 tinguished through their insolubility in fat solvents, digestion 

 in artificial gastric juice, and postive xanthoproteic and nin- 

 hydrin reactions. 



(2) WW-STt; PRODUCTS 



Inorganic Sphaerocnistalf;. Reddish-brown, weakly bi-re- 

 fringent sphaerocrystals occur in the intestine of many para- 

 sitic nematodes, including Asearis, Camallaniis, Strongylns, 

 Ancylostoma, and Trieliiiris. These structures are similar in 

 appearance to rhabditin both in transmitted light and between 

 crossed Nichols, although in totomount preparations and sec- 

 tions they are not liirefringent. The optical activity in this 

 case can only be observed when the crystals are isolated. Un- 

 like rhabditin, they are dark in dark field illumination and 

 are insoluble in water, acetic acid, NaOH (all concentrations) 

 and saliva. They are also insoluble in alcohol and xylol and 

 are not affected by gastric or jiancreatic enzymes. Askanazy 

 (1896), Looss (1905) and Faure-Fremiet (19r2) regarded 

 them as products of haenu)globin resorption, while Lievre 

 (1934) was unable to establish the presence of haemoglobin 

 in the intestine of Ascaris lumbricoidex and Parascaris 

 eqworum by spectroscopic analyses. He demonstrated the pres- 

 ence of haemoglobin by this means in 7.)% of the specimens 

 of Toxocara cants examined. A'on Kenmitz (1912) identified 

 them as zymogen granules and Quack (1913) identified them 

 as gypsum (CaS04.2II,0 ). The writers find that these crystals 

 may be obtained relatively- pure by boiling the intestine in 

 10% KOH and wa.shing in a centrifuge. Crystals prepared 

 in this manner are not charred by heating on a gla.ss slide to 

 the melting point of glass. They may be dissolved liy heating 

 in concentrated HOI and when cooled recrystallize in the gen- 

 eral habit of CaS04.2H=0. Such crystals are birefringent and 

 obliquely extinct. This is presumptive evidence that they are 

 gypsum. However, a reddish-brown residue is left when the 

 slide is dried. This residue stains blue in dilute HCl-potas- 

 sium ferrocyanide, indicating the presence of iron; direct ex- 

 periments on the sphaerocrystals jiroduces the same result in 

 partially dissolved ( swollen 1 crystals. Some compound con- 

 taining iron is evidently present as an adsorption within the 

 sjihaerocrystals. As evidence of the association of sjihaerocrys- 

 tals with a blood feeding mode of life, Tornquist (1930) point- 

 ed out that ('aiiKiUaniix. which is known to feed on blood, has 

 them, while Ciiciithiini.i. wliicli does not feed on blood, does not 

 possess them. One might add that adult oxyurids and thelasto- 

 matids, as well as the first three larval stages of Strongylns. and 

 Camallanus are also devoid of them. The evidence is entirely 

 circumstantial. The occurrence of grossly similar insoluble 

 sphaeroids in the intestine of Theristus setosus (see Olivaceous 

 sphaeroids) and other free-living nematodes casts some doubt 



on the above interjuetation since they also contain iron. Lievre 

 (1934) interprets positive tests for iron in the intestine of 

 Asearis as due to substances obtained from animal and vege 

 table food, not haemoglobin. 



Olivaccovs sphaeroids. Reddish brown, apparently non-bire 

 fringent sphaeroids were observed by the writers (1938) in the 

 intestinal cells of Tlieristus setosus (Fig. lOOB). They have 

 the following characteristics: Not blackened by osmic acid; 

 not colored by Scharlach R, insoluble in alcohol; ninhydrin 

 and xanthoproteic reactions negative; blue in neutral violet; 

 blue in crystal violet, blue in nile-blue sulphate; not digested 

 by artificial gastric juice or diastase; soluble in 10% HCl and 

 2% KOH but not in ^0% acetic acid or 2% HCl; blue in 

 potassium ferricyanide followed by 1% HCl. From these ob- 

 servations it seems that the sphaeroids must consist of an or- 

 ganic ferrous iron salt or a salt of a weak base (Fe(OH)«) and 

 a weak acid. Similar sphaeroids were also observed in Dory- 

 laimtis stagnatis and an unidentified oncholaimid and Diptos- 

 capter cnronata (Fig. lOOD). 



Crystals. Stefanski (1916) and Cobb (1918) observed poly- 

 hedral colorless birefringent crystals in the intestine of Ironus. 

 They are very similar in appearance (Fig. lOoP) to triple 

 phosjihate, and, according to Stefanski, they are very soluble 

 in acetic acid and potassium hydroxide; slightly soluble in 

 hot water and insoluble in cold water, alcohol, ether, chloro- 

 form, and acetone; are not stained by iodine-potassium-iodide, 

 but stain with eosin and fuchsin. Isolated colorless polyhedral 

 crystals have also been observed in Tripyla (Fig. lO.^A). 



Crystal aggregates in mermithids were observed bv Meiss- 

 ner (18.53), Rauther (1906), Ilagmeier (1912) and'christie 

 (1936). These are first seen in vacuoles of the post nodal re- 

 gion of preparasitic larval Agamermis deeaiidata. Their num- 

 ber increases with age and in old adult specimens similar 

 crystals have been observed in the body cavity. They are very 

 similar in appearance (Fig. 107D) to uric acid and allantoin; 

 are birefringent, obliquely extinct; insoluble in water, alcohol, 

 ether, 10% ammonium hydroxide, 10% acetic acid, 10% HCl, 

 glycerin, and V^ -saturated ammonium sulphate. Presumably 

 they represent a nitrogenous product. 



In niploscapter comnata large (piadrate tablets, colorless to 

 yellowish brown, have been observed by the writers (Fig. 

 lOOD). Like oliv.aceous sphaeroids, they are non-birefringent, 

 soluble in 2% sodium hydroxide and 10% hydrochloric acid 

 but are insoluble in 1% HCl, alcohol and glycerin. They give 

 a negative ninhydrin reaction and a positive potassium fer- 

 ricyanide — 1% HCl reaction. Therefore they appear to be a 

 ferrous iron compcjund, probabl.v organic in nature. It is 

 possible that they may be crystals of the same substance com- 

 posing the olivaceous sphaeroids. 



(3) JnSCEI.L.\NEOUS "URANULES" 



In the majority of instances the cell contents of the nema- 

 tode intestine have not been studied chemically. The term 

 "granule" is, of course, chemically meaningless. Since at 

 least four distinct substances are known to exist in a sphaeroi- 

 dal state, namely, rhabditin, fat, protein, gypsum and oliva- 

 ceous sphaeroids each form must be considered with care. 

 With living specimens, dark field illumination is sufficient to 

 separate the globules of fat and protein from the sphaerocrys- 

 tals of rhabditin and gypsum. In addition, the first two are 

 colorless while the last three are yellowish brown to reddish 

 brown. However, still other types may be discovered. Non- 

 birefringent brownish or yellowish refractive "granules" are 

 present in the intestinal epithelium of Mctnncholaimtis, Si- 

 phonolainiits, Ironus, and otlier forms. Such "granules" are 

 strongly refractive and non staining (? olivaceous sphaeroids). 

 In addition one finds moderately refractive basophilic "gran- 

 ules" in the intestine of such forms as Dori/laimopsis, and 

 Plectiis, and special cells of the intestine of Synonchirtla. 

 Cobb (1922) has described birefringent sphaerocrystals in 

 s])ecial cells of Ei(rystomina as " marionellin " and we find 

 these to be relatively insoluble and basophilic. In other forms 

 acidophilic "granules" have been observed. For morphologi- 



Fii; 



lOI!. INTHSTINE IX AiSCAKIDIN A, SIMRITRIXA AND OAMALLANINA 



A — Rhiyoneina ittfecta. B-C — Spirnnoura nffine, (B — cross; C — 

 longitudinal in prcrectal rcRioii showing parasites). D — Heterakin 

 iiaUinae. E-F — Oxjntrit etiui (E — superficial: P — longitudinal). G-H 

 — Macruris fiinnlii/nlfrn (<t — cross; H — supei-iicial) . I — Anisakinid 

 internal view showing ridges as seen in dissection .T-Q — Aticaris 

 InmhrietyidfH (.1 — noimal; 1. bacillary layer; 2. subbacillary layer; 'A. 

 plasma cap: 4. endophtsni; .5. jilasnia string; fi. basioplnsni ; 7. basal 

 lamella.; K — liinucleate cell, one nucleus degenerating; L — early cy- 

 tnyilasmic degeneration: M — lale cytolplasniic degeneration; N — nor- 

 mal nucleus; (> — early degenerating nucleus; T* — mid degenerating 

 nucleus; Q late (le'.;eneriitiMg nucleus). R S Tnjorfirti rtinin ( R— 



intestinal cell with Ca.ial silver; S — with Iodine vapor, glc. glycogen, 

 grn i insoluble gran\ile (sphaerocrystal) T-U — (Inaihostoma spini- 

 (ji'nini V — liirtutarifi I'nlnradiensiti (grn b basophilic granule). W- 

 V — Pliihinietra rubra ( W — surface view; X — ventricular region; Y- — ■ 

 uiidregion ) . Z — Phf/.^nJoplera retujta (1. Bacillary layer; 2. .sub- 

 bacillary layer; 3. plasma cap; 4. endoplasm ; 5. plasma string; 6. 

 basal lamella). AA — Tnnqvfi tiara. BB — A,scaropliis (CystidirnJa 

 liarivoadi) . TC — Camattanas atnerirarxan. K-F, after Martini. 1916, 

 Ztschr. Wiss. Zool. v. 110; I, after Cobb. 1SK8, Heilraege 7,ur Anat- 

 oniie und Ontogenie dec Nenuxtoden ; R-.S, after .\rgeseanu, 19.')4. 

 Crnnpt, Rend. S«ic. Biol, v. 110; remainder original. 



104 



