A Sarcina bacterium linked to lethal disease in sanctuary chimpanzees in Sierra Leone.

Human and animal infections with bacteria of the genus Sarcina (family Clostridiaceae) are associated with gastric dilation and emphysematous gastritis. However, the potential roles of sarcinae as commensals or pathogens remain unclear. Here, we investigate a lethal disease of unknown etiology that affects sanctuary chimpanzees (Pan troglodytes verus) in Sierra Leone. The disease, which we have named "epizootic neurologic and gastroenteric syndrome" (ENGS), is characterized by neurologic and gastrointestinal signs and results in death of the animals, even after medical treatment. Using a case-control study design, we show that ENGS is strongly associated with Sarcina infection. The microorganism is distinct from Sarcina ventriculi and other known members of its genus, based on bacterial morphology and growth characteristics. Whole-genome sequencing confirms this distinction and reveals the presence of genetic features that may account for the unusual virulence of the bacterium. Therefore, we propose that this organism be considered the representative of a new species, named "Candidatus Sarcina troglodytae". Our results suggest that a heretofore unrecognized complex of related sarcinae likely exists, some of which may be highly virulent. However, the potential role of "Ca. S. troglodytae" in the etiology of ENGS, alone or in combination with other factors, remains a topic for future research.

Priority of the genus name Clostridium Prazmowski 1880 (Approved Lists 1980) vs Sarcina Goodsir 1842 (Approved Lists 1980) and the creation of the illegitimate combinations Clostridium maximum (Lindner 1888) Lawson and Rainey 2016 and Clostridium ventriculi (Goodsir 1842) Lawson and Rainey 2016 that may not be used.

In a recent publication that attempts to deal with the growing problem of taxa being added to the genus Clostridium that are outside of Clostridium (16S rRNA) group I, a solution is proposed that seeks to limit the genus Clostridium Prazmowski 1880 (Approved Lists 1980) to a small number of species 'related' to the type species, Clostridium butyricum Prazmowski 1880 (Approved Lists 1980). It has been proposed that this genus should also include members of the genus Sarcina Goodsir 1842 (Approved Lists 1980), Sarcinamaxima Lindner 1888 (Approved Lists 1980) and Sarcinaventriculi Goodsir 1842 (Approved Lists 1980), the latter being the nomenclatural type of the genus Sarcina Goodsir 1842 (Approved Lists 1980). In making proposals to treat the genus name Sarcina Goodsir 1842 (Approved Lists 1980) as a synonym of ClostridiumPrazmowski 1880 (Approved Lists 1980), reference is made to the wording of the International Code of Nomenclature of Bacteria. However, while that wording is factually correct, other parts of the Code are relevant to this issue and clearly indicate that the proposed course of action is not sanctioned by texts that have not been directly made reference to. Rather than avoiding confusion it has been contributed to, and it is necessary to document where the problems lie.

Sarcina organisms in the gastrointestinal tract: a clinicopathologic and molecular study.

Sarcina organisms were first observed in and recorded from the stomach contents of a patient suffering from vomiting by John Goodsir in 1842. Since that time, their fine structure, phylogenetic classification, and biochemical characteristics have been described. Although numerous cases of fatal disease have been attributed to this organism in the veterinary literature, only a few human cases have been documented. As a result, whether this organism causes disease in humans has not been definitively established. We report the clinicopathologic findings in a series of 5 patients with Sarcina-like organisms identified in upper gastrointestinal endoscopic biopsies with molecular confirmation. Our findings have shown that the organism is most commonly found in patients with a history of gastric outlet obstruction or delayed gastric emptying. Although many of the patients do not demonstrate direct mucosal injury from the organism, the presence of a concurrent gastric ulcer puts the patient at increased risk for complications such as emphysematous gastritis or perforation. The finding of Sarcina organisms should prompt further investigation for functional causes of gastric outlet obstruction and delayed gastric emptying, such as occult malignancy.

Phylogenetic placement of Sarcina ventriculi and Sarcina maxima within group I Clostridium, a possible problem for future revision of the genus Clostridium. Request for an opinion.

The 16S rRNA gene sequences of Sarcina ventriculi DSM 286T (T = type strain) and Sarcina maxima DSM 316T were determined. Phylogenetic analysis revealed that these two species are closely related to each other and belong to group I Clostridium (sensu Johnson and Francis). The implications of these phylogenetic findings for future revision of the genus Clostridium are discussed.

A separation of staphylococci and micrococci based on serological reactivity with antiserum specific for polyglycerophosphate.

A serological reaction with the antiserum against heterophile polyglycerophosphate (PGP) was evaluated for genus level differentiation among strains of Staphylococcus and Micrococcus-Sarcina spp.. Hot saline extracts from whole cells of Staphylococcus spp. strongly reacted with the PGP antiserum, whereas those of Micrococcus-Sarcina spp. did not. Likewise, phenol-water extracts from whole cells of Micrococcus-Sarcina spp. were not reactive with the PGP antiserum, although the extracts of staphylococcal cells again gave a strong reaction with the antiserum. This study indicates that extracts from Micrococcus-Sarcina spp. have no antigen reactive with the PGP antiserum and can thus be differentiated from extracts of Staphylococcus spp. which react strongly with the PGP antiserum.

Classification of micrococci on the basis of deoxyribonucleic acid homology.

The DNA homology relationships of 25 micrococci (15 strains of Micrococcus, eight strains of Sarcina and two strains of Staphylococcus) were studied by the deoxyribonucleic acid hybridization method using nuclease S1, an endonuclease specific for single-stranded DNA molecules. Nineteen of the strains were classified into three groups. Group I contained Micrococcus lysodeikticus IAMI056, M. luteus IAMI1010, M. flavus IAMI2005 and IAMI2006, Sarcina flava IAMI2007 and IAMI1006. S. subflava IAMI2009, S. lutea ATCC381, and ATCC382, and M. luteus IAMI1006. Group II contained M. roseus IAMI315, ATCC412, ATCC185 and IAMI295. Group III contained S. lutea IAMI099, IFO3232 and ATCC383, M. varians ATCC399 and Staphylococcus lactis ATCC15306. Micrococcus luteus IAMI097, M. varians ATCC19099 and ATCC19100, M. conglomeratus IAMI459 and IAMI470, and St. aureus IAMI011 could not be assigned to any of the three groups. The grouping corresponds to that derived from the results of differential lysis by lysozyme, 'lytic enzyme 2' from Cytophaga sp., or Streptomyces albus G enzyme; and to types of peptidoglycan in the cell walls and genetic transformation. The usefulness of classification based on sensitivity to various lytic enzymes was demonstrated. Group I probably coincides with M. luteus of Bergey's Manual of Determinative Bacteriology (1974), and groups II and III with M. roseus and M. varians respectively.

Use of antibody to membrane adenosine triphosphatase in the study of bacterial relatioships.

An antiserum to Ca(2+)-activated adenosine triphosphatase from membranes of Micrococcus lysodeikticus cross-reacted in agar gels with membrane adenosine triphosphatases from other pigmented micrococci and related species. Species of Micrococcus and Sarcina showed different levels of inhibition of adenosine triphosphatase activities in heterologous reactions with antiserum. Inter- and intraspecific relationships based on the inhibition reaction were compared with an independent parameter, namely the quantitative and qualitative composition of the bacterial membrane phospholipids and fatty acids. The guanine plus cytosine contents in the deoxyribonucleic acid of the species studied correlated well with the serological cross-reactivity of adenosine triphosphatases from their membranes. The types of cross-bridges found in the peptidoglycans of these cocci were also compared with the other properties. The results suggest that an antiserum specific for a major membrane protein may be a reliable and most useful adjunct in studying bacterial serotaxonomy.

Comparative allostery of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthetase as a molecular basis for classification.

The allosteric pattern of control for 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthetase has previously been shown to be strongly conserved among the member species of a given genus in bacteria. The implications of this finding as a procedural tool of bacterial phylogeny were pursued by a study of two organisms, Sporosarcina ureae and Aeromonas formicans, the taxonomic positions of which have been historically controversial. S. ureae has characteristics of both Bacillaceae and Micrococcaceae, and A. formicans has characteristics of both Enterobacteriaceae and pseudomonads. Since the patterns of control for DAHP synthetase in all four of these microbial groups are different from one another but internally homogeneous within each group, the results obtained from the two test organisms were unambiguous. It was concluded that S. ureae is properly classified within Bacillaceae, probably deserving generic rank, and that A. formicans belongs with the family Enterobacteriaceae.

Genetic relationship between Sarcina ureae and members of the genus Bacillus.

Deoxyribonucleic acid-ribonucleic acid (DNA-RNA) and DNA-DNA hybridization studies were performed to determine the degree of genetic relatedness between Sarcina ureae and several members of the genus Bacillus. DNA-RNA hybridization showed a high degree of homology between S. ureae RNA and DNA from Bacillus species having a similar guanine plus cytosine content. The DNA from other genera of the family Micrococcaceae showed less homology with S. ureae RNA than did that of the Bacillus species tested; however, this homology was not found between the DNA of S. ureae and DNA from these Bacillus species or DNA from the other Micrococcaceae tested. Transformation with Bacillus DNA, infection with representatives from several major classes of Bacillus phages, and electrophoretic analysis of proteins in crude extracts of these strains were also attempted as a further test of the genetic relationship between the genera. These experiments did not support the belief that the two groups are closely related genetically.

Identification of Micrococcaceae in clinical bacteriology.

The cellular morphology, identifying physiological characteristics, and a key to the human genera of Micrococcaceae are presented with flow charts for identification of aerobic and anaerobic isolates. These flow charts can be amended as desired, depending upon the degree of accuracy desired. Micrococcaceae isolates in a 350-bed private general hospital during a 15-week period are tabulated to show relative numbers of the different genera and species, with their probable relationship to infection or contamination. Only 11 of the 220 Micrococcaceae isolates were not Staphylococcus; no Sarcina or Peptococcus were isolated. Of the Staphylococcus isolates, 61% were S. epidermidis. Almost 18% of the S. aureus isolates were coagulase-negative. Of the S. aureus isolates, 80% of the coagulase-positive isolates were infecting agents, as were 67% of the coagulase-negative S. aureus isolates, compared to only 48% of S. epidermidis isolates. Two of four Gaffkya isolates but only one of seven Micrococcus isolates were infecting agents. If coagulase production is used as the sole criterion for speciation of staphylococci, and Micrococcus is not differentiated from Staphylococcus, the term "coagulase-negative staphylococci" does not differentiate three distinct levels of pathogenicity. Coagulase-negative S. aureus is more virulent than S. epidermidis or Gaffkya, which are more virulent than Micrococcus or Sarcina.

Taxonomy of psychrophilic strains of Bacillus.

The morphological and physiological characteristics of 20 isolates of psychrophilic Bacillus were compared with 29 strains representing nine species of mesophilic Bacillus and 2 strains of Sporosarcina ureae to determine the taxonomic position of the psychrophiles. The psychrophiles formed four distinct groups which were sufficiently different from the mesophiles to warrant their designation as new species of Bacillus. The names B. psychrosaccharolyticus, B. insolitus, B. globisporus, and B. psychrophilus are proposed for the new species.