Ten varieties of North American shellfish were examined for the occurrence of Cristispira in their styles. A cristispira was found in various numbers in Ostrea virginiana, Venus mercenaria, and Modiola modiolus, but none in Ensis americana, Mya arenaria, Mactra solidissima, Pecten irradians, Mytilus edulis, Fulgur canaliculatus, or Nassa obsoleta. Of 298 oysters, only 128 showed the crystalline styles, in which cristispiras were present in 99. Active cristispiras were found in 59 styles only and degenerated forms in the remaining 40. In 110 clams (Venus mercenaria) 70 styles were found, and only 8 of these contained cristispiras; 5 yielded active and the other 3 degenerated cristispiras. In 97 modiolas there were 73 styles, only 4 of which contained cristispiras.
The physical properties of the crystalline styles of these shellfish varied considerably. The styles of the oysters were moderately soft, and when exposed to the air or mixed with sea water they underwent liquefaction, forming a clear, viscid material. The styles from clams and modiolas were opaque and were more firm, not easily crushed even in a mortar. The styles of the scallops were the most solid of all the styles examined. It happened that the softer the styles, the more frequent was the occurrence of the cristispira; in fact, no cristispira was detected in styles other than those of oysters, clams, and modiolas, of which oysters had the softest styles and the largest percentage of cristispira invasion.
The following observations were made regarding the structure of the cristispira found in oysters. The body is a long, flexible cylinder, with blunt extremities, towards which the diameter gradually diminishes. In motion the body rapidly stretches and contracts, forming in the contracted state several serpentine undulations. A membranous appendage (Gross' crista) winds about the body throughout its entire length. The inner margin is in connection with the body, the outer margin is free and is distinctly heavier. The latter is undulatory; that is, the width of the membrane, or crista, is narrower at some points than at others. The membrane is composed of numerous fine fibrils running in a roughly parallel or slightly oblique course, showing interwoven narrow meshes; at the outer margin there is a dense smooth ridge.
The contour of the body is highly refractive, as if possessing a cell membrane. The interior structure, as revealed by dark-field illumination, is an almost homogeneous, less refractive substance, but there are present minute highly refractive granules more or less symmetrically arranged. There is no definite cross-bar or chambered structure. On the other hand, when vital staining with brilliant cresyl blue is applied, there appear numerous paired masses of lavender hue at fairly regular intervals, suggesting the cross-bar aspect of a stained specimen. In a few specimens there was seen a dim outline of cross-bar effect. Neutral red, Bismarck brown, and crystal violet all bring out deeply stained granules and reticular structure but no definite cross-bars.
The crista is a fibrillar structure, connected with the body at its inner edge. The outer margin is a thickened bundle of fibrils running an undulating course along the entire length of the crista. The crista is elastic and when detached from degenerated organisms assumes a rather regularly wound spiral, consisting of longitudinal bundles of fibrils (Figs. 36 to 38). A fragment of two or three waves may be encountered in a preparation containing many degenerated organisms (Fig. 39). The composition of the crista can best be studied in degenerated remains of the organism. During the life of Cristispira it is stretched or relaxed according to the contraction or extension of the body. The elasticity of the crista appears to furnish the organism with a propelling and rotating power upon its extension after being drawn tightly to the body by some contractile apparatus (myoneme) present somewhere within the cell body. The crista serves as a rudder and propeller for the swimming organism. It is interesting to compare here the elastic and regularly waved flagella of certain bacteria and spirochetes; it is possible that the crista of Cristispira is a highly modified form of flagella.
The nature of the substance which stains dark blue with Giemsa's stain is not known, but it does not give a chromatin reaction. By Heidenhain's iron-hematoxylin method it takes a dark grayish tint, similar to the cell wall or crista, which are also dark gray. This substance was regarded by Gross and Zuelzer as volutin, which is of nutritive origin. It is probable that there are also embedded within it minute chromidial elements. Multiplication is by transverse fission.
Cristispira balbianii is parasitic and does not survive more than a few days in ordinary sea water emulsion, even at its optimum temperature. In its natural habitat, or the crystalline style, it is usually pure, but is sometimes found in association with a tiny spiral organism (Spirillum ostreæ). The cristispiras in the styles seem to diminish rapidly when oysters are collected from their beds and transferred elsewhere; oysters kept in tanks or cars for several days do not contain the cristispiras, and in opened oysters the styles disappear promptly at room temperature.
All efforts to cultivate this organism have failed.