Clinical application of novel sample processing technology for the identification of salmonellae by using DNA probes.

Two hundred and fifty clinical fecal specimens collected over a 7-month period were analyzed for the presence of salmonellae by a rapid DNA hybridization procedure. Hybridizations were performed by using a novel specimen processing protocol called wicking and a previously unreported 1,600-base-pair probe cloned from Salmonella enteritidis DNA. The probe was shown to be reactive with all 70 Salmonella serotypes tested and not reactive with 101 stock strains of other enteric bacteria. Southern analysis of 30 Salmonella isolates representing 22 serotypes suggested that the probe sequence was highly conserved, appearing as a 1,600-base-pair band in a BglII digest of isolate DNA in 29 of 30 isolates and as a 2,300-base-pair fragment in 1 of the isolates. The probe correctly identified all salmonellae (nine isolates) among 47 H2S-producing colonies tested from among 250 clinical specimens cultured on xylose-lysine-desoxycholate medium. Salmonellae grown on xylose-lysine-desoxycholate medium gave consistently higher hybridization values than did those grown on either MacConkey or Hektoen enteric agar. In addition, of eight gram-negative broth enrichments in which salmonellae were identified by conventional means, seven were probe positive. The use of this nucleic acid probe and hybridization technique provides a simple and rapid identification of Salmonella species.

Cephalic flexure formation in the chick embryo.

The cephalic flexure, found in all vertebrate brains, is a ventrally directed bend through the mesencephalon, and a ventral bulging and elongation of the prosencephalon. Most sources say the cephalic flexure is caused by differential growth. We have measured the changing angle of flexure through time and find that flexure occurs between chick embryo stages 10 to 15. We measured, during these stages, the lengths, thicknesses, and volumes of the floor and roof of the mesencephalon and of the prosencephalon. As expected, during flexure the mesencephalic roof elongates much more than the floor. Both roof and floor increase in thickness, and mesencephalic roof volume increases twice as much as floor volume. However, prosencephalon, which does not bend, also has differential growth between roof and floor, but the growth is taken up in complex changes of shape other than flexure. There are sufficient numbers of mitoses in the brain to account for the observed tissue growth, assuming accompanying cell enlargement. We deleted brain parts adjacent to the mesencephalon before flexure and the mesencephalon bent, so migration of cells from or to these adjacent parts to contribute to the differential growth of the mesencephalon is unlikely. We reduced cerebrospinal fluid pressure during flexure by explanting heads to the chorioallantoic membrane, or into New cultures. The mesencephalon of explanted heads bends, but the prosencephalon fails to elongate. We conclude that differential growth may be necessary for mesencephalic flexure in the chick embryo, but other factors that decide the disposition of the products of growth in space must determine the shape.