A monoclonal antibody reacting with the cell envelope of spirochaetes isolated from cases of intestinal spirochaetosis in pigs and humans.

A monoclonal antibody (mAb) designed BJL/AC1 was prepared against the cell envelope of an intestinal spirochaete (strain 3295) that was isolated from a pig with intestinal spirochaetosis. The mAb reacted with a band of approximately 29 kDa in cell envelope preparations from 13 porcine and 11 human spirochaetes isolated from cases of intestinal spirochaetosis, but did not react with preparations made from a range of other intestinal spirochaetes. Immunogold labelling demonstrated that the reactive epitope was located on the cell envelope of the strains causing intestinal spirochaetosis. The mAb was used in an indirect immunofluorescence test to detect spirochaetes in the faeces of pigs with experimentally induced intestinal spirochaetosis. The mAb should prove to be a useful reagent for detection and identification of spirochaetes that are specifically associated with intestinal spirochaetosis.

The porcine intestinal spirochaetes: identification of new genetic groups.

The weakly beta-haemolytic isolates were divided into 56 electrophoretic types (ETs), contained in three distinct genetic groups (A,B and C). Group A corresponded to the genus Serpulina, and could be divided into three divisions. It contained 17 weakly haemolytic isolates in divisions b and c, as well as all 98 isolates of S. hyodysenteriae, located in division a. All seven weakly beta-haemolytic isolates that produced indole and had alpha-glucosidase but not alpha-galactosidase activity fell into division b. These spirochaetes may represent a distinct species. The other ten weakly beta-haemolytic spirochaetes, in division c, fitted the description of S. innocens. Group B contained 17 of the weakly beta-haemolytic isolates (18.9%) in ten ETs. Isolates in this group differed from typical S. innocens in that they lacked alpha-galactosidase activity. Group B represented a distinct group of weekly beta-haemolytic spirochaetes, which may constitute a new genus. Group C contained 56 of the weakly beta-haemolytic isolates (62.2%) located in 29 ETs. The original isolate from "spirochaetal diarrhoea" (P43/6/78-Taylor et al., 1980) was located in this group, together with Australian isolates from a similar condition. Spirochaetes in group C were morphologically distinct from those in groups A and B in that they possessed only four, five or occasionally six, subterminal axial filaments, were more slender, and had more pointed ends to their cells. We consider that group C represents a new genus of spirochaetes, members of which may be associated with spirochaetal diarrhoea.

Cristispira from oyster styles: complex morphology of large symbiotic spirochetes.

Crystalline styles (digestive organs) of bivalve mollusks provide the habitat for highly motile bacteria. Styles from freshly-collected oysters, Crassostrea virginica, were studied by electron microscopy; Cristispira spirochetes were abundant in these organs. Detailed study reveals these spirochetes to be among the most complex prokaryotic cells known. More than 600 periplasmic flagella and an adhering outer lipoprotein membrane (e.g., a 270 degrees sillon) form the ultrastructural basis for the "crista," first described by light microscopy. Unique rosette structures corresponding to the "chambers" or "ovoid inclusions" of light microscopy were detected at the periphery of all protoplasmic cylinders. Polar organelles and linearly aligned flagellar insertions are conspicuous. In size and complexity, Cristispira more resembles Pillotina, Diplocalyx, Clevelandina and Hollandina (large spirochetes symbiotic in termites) than it does Treponema. Cristispira pectinis (Gross, 1910), the type species; Spirillum ostrea (Noguchi, 1921); and another, less frequent bacterial symbiont are the predominant inhabitants of the dense style matrix. The ultrastructure of the spirillum and an electron micrograph of the third bacterium are shown.

Infection of the colon of the rhesus monkey by spiral-shaped organisms.

Intestinal spirochetosis, an infection of the mucosa by spiral-shaped organisms, was studied in clinically normal rhesus monkeys (Macaca mulatta) by histology, transmission and scanning electron microscopy. The incidence of intestinal spirochetosis was 42% in 221 monkeys. Spiral organisms stained with hematoxylin and eosin (HE) appeared as a broad basophilic haze on the colonic surface and were strongly positive by the Warthin-Starry stain. Spiral-shaped bacteria include two structurally different organisms: spirochetes and flagellated microbes. They intimately populated the brush border of the surface of the epithelium of the large intestine. They were absent in the crypts and in the small intestine. Infection by spirochetes produced no alteration of cytocomponents of the underlying host structures. Spirochetes and flagellates infrequently penetrated beyond the brush border into the epithelial cytoplasm and also into the lamina propria. Even in cases where invasion was documented, no inflammatory response was found.

Small pectinolytic spirochetes from the rumen.

Two strains of small spirochetes utilizing pectin as sole source of energy for growth were isolated from the rumen of cattle and partially characterized. The pectinolytic enzyme produced by one of the organisms was classified as a poly-1,4-alpha-D-galacturonide lyase (EC 4.2.2.2). A pectinesterase (EC 3.1.1.11) was also produced.

Possible evolutionary significance of spirochaetes.

Large symbiotic spirochaetes of the family Pillotaceae (e.g. pillotinas) are found in dry wood and subterranean termites (Hollande & Garagozlou 1967). These morphologically distinctive spirochaetes comprise several genera. Some of them contain microtubules within their protoplasmic cylinders. They demonstrate a variety of relations with their termite and protist hosts. Some are free-living within the lumen of the intestine, some tend to be associated with filamentous and other bacteria, some are found regularly coursing between the numerous undulipodia ( = eukaryotic flagella, cilia, and other (9 + 2) organelles of motility) of hypermastigotes and polymastigotes. Still other smaller termite spirochaetes are regularly attached to protists via specialized attachment sites. Some even form motility symbiosis with their host protists. The analogy between the behaviour of host-associated spirochaetes and the possible steps in the origin of the undulipodia and mitotic system of eukaryotes is discussed briefly.

Pillotinas and hollandinas: distribution and behaviour of large spirochaetes symbiotic in termites.

Pillotina spirochaetes have been observed in the hindguts of wood-eating cockroaches (Cryptocercus punctulatus), and in 25 out of 28 species of termites examined. They were especially abundant in 21 species of dry wood termites of the family Kalotermitidae, from Europe, North America and Australia. These included many species of Kalotermes and one or a few of the following: Glyptotermes, Bifidotermes, Neotermes, Ceratokalotermes, Paraneotermes, Cryptotermes, Porotermes, Marginitermes, Pterotermes, Zootermopsis, Reticulitermes, Coptotermes, Heterotermes, and nasutitermitids. Identifications of pillotinas were made on the basis of large size (0.5--2 micromtere in diameter, 50 to greater than 100 micrometers in length) and wave pattern; these were verified by electron microscopy in K. schwarzi, Pterotermes occidentis and others. Pillotinas were also present in all species of subterranean termites (Family Rhinotermitidae) examined, and in the most primitive Australian termite, Mastotermes darwiniensis (Family Mastotermitidae). They were not observed in damp wood termites (Family Hodotermidiae). Pillotinas are invariably associated with a rich, complex xylophagous microbial community composed primarily of motile prokaryotes, and hypermastigote and polymastigote flagellates. Some have been previously described by those primarily concerned with termite hindgut protozoa. Observations were made on their modes of behaviour, division, and microbial associates. A new genus of spirochaetes, Hollandina, is also described. It is distinguished from Pillotina by a smaller size and several ultrastructural features, but is otherwise closely related taxonomically. Evidence is provided to support Hollande and Gharagozlou's (1967) concept that the pillotinas and hollandinas deserve the taxonomic status of 'family' and that they should be classified with the cristispire siprochaetes a-cording to the scheme developed by Hovind-Hougen (1976). Spirochaetes are treated as a Phylum of the Kingdom Monera (Prokaryota) in the five kingdom system of Whittaker (1969).