Evaluation of the Modified R/B System for Identification of Enterobacteriaceae

In comparison with conventional procedures, a modified R/B Enteric Differential System permitted more accurate identification of enteric bacteria than did the original R/B system.     

Evaluation of the Auxotab Enteric 1 System for Identification of Enterobacteriaceae

An evaluation of the accuracy and convenience of the Auxotab Enteric 1 System for identification of Enterobacteriaceae was performed with 160 bacteria. Identification at the species level was correct in 134 (83.8%) instances and at the generic level in 144 (90%) instances. Sixty strains failed to achieve the minimal concentration of organisms required to complete the identification process within 7 hr. The system was judged to be laborious and to present a potential hazard to those working with it.     

Comparison of Conventional Methods, the R/B System, and Modified R/B System as Guides to the Major Divisions of Enterobacteriaceae

Primary grouping of more than 2,000 members of Enterobacteriaceae, freshly isolated from clinical specimens, was performed on the basis of eight conventional laboratory tests used routinely in this laboratory and by the R/B system. Results indicate that both systems perform well in this first step in the identification of various medically significant species of the family. Difficulties encountered with certain reactions of the R/B system were corrected with a physical modification of the tubes. An additional 100 representatives of Enterobacteriaceae were compared with conventional methods and the modified R/B system. Results indicate improvement in observing and interpreting findings with the R/B system.     

Evaluation of the Minitek System for Identification of Enterobacteriaceae

Clinical isolates (869) and stock cultures (35) of Enterobacteriaceae were tested in parallel with the Minitek and conventional systems. The Minitek correctly identified 822 of 904 cultures. When a deoxyribonuclease plate was inoculated along with the Minitek, it was possible to speciate Enterobacteriaceae within 24 h. False-positive hydrogen sulfide reactions were the major fault with this system. Reactions were clear-cut and easy for technologists to read.     

API System: a Multitube Micromethod for Identification of Enterobacteriaceae

The API system for identification of Enterobacteriaceae was evaluated with 366 cultures. Overall accuracy of identification was 96.4%; of the 13 cultures misidentified, 7 were atypical strains.     

Evaluation of Accuracy of Multitest Micromethod System for Identification of Enterobacteriaceae

The Analytab system of 20 biochemical tests for identification of Enterobacteriaceae was evaluated in parallel with conventional tests on 128 Enterobacteriaceae, 5 Aeromonas, and 1 Yersinia enterocolitica. The results of tests for H(2)S and indole production, citrate utilization, lysine and ornithine decarboxylase, arginine dihydrolase, nitrate reduction, beta-galactosidase, and fermentation of arabinose, rhamnose, mannitol, and glucose showed almost complete agreement between the two systems. Eighty-eight per cent of Enterobacteriaceae were correctly speciated with the Analytab system; on repeat testing with heavier inocula of organisms failing to ferment glucose initially, the proportion of Enterobacteriaceae correctly speciated became 93%.     

Evaluation of the BBL Minitek System for the Identification of Enterobacteriaceae

Thirty-one different substrate disks were tested in parallel with comparable, prepared media (BBL) against a minimum of 300 cultures of Enterobacteriaceae. An overall correlation of 98% was observed with all the disks tested. In addition, the system was used to identify 461 fresh isolates of Enterobacteriaceae in parallel with conventional media using the schema used at the Veterans Administration Hospital, Baltimore. An overall correlation of 97% was observed. Minitek is a time and space saving system. It is accurate and easily adapted to the clinical laboratory. A wide variety of substrates are available, allowing most laboratories to use their own schema. The long shelf life of most disks is a definite advantage.     

一个1B/1R小麦-黑麦染色体易位的鉴定

本研究对冬小麦品系73(36)9-1的1B/1R易位染色体进行了遗传分析。发现73(36)9-1有一对随体染色体,它的两个亲本矮丰四号及洛夫林10(Lovrin lo)分别有两对和一对随体染色体。观察用矮丰四号回交的F_1,绝大部分花粉母细胞的中期染色体都能正常配对,而用洛夫林10回交的,除了多数产生两个单价体之外,正常配对的情况也能经常看到。同时还发现,73(36)9-1和“中国春”双端体(CSDT)的1BL能很好地配对并形成一个棒状的异形二价体,而它和CSDT 1BS的染色体则主要产生20″+1′+t′的构型,从而证明易位发生在1B染色体的短臂,并且该易位的片段来自黑麦染色体1RL。本文还讨论了该易位发生的可能途径,推断是由于在F_1花粉母细胞中的两个单价体(一个是小麦染色体1B,一个是黑麦染色体1R)同时进行错分裂之后产生的两种端着丝点染色体(1BL和1RL)重新并合形成的,因而冬小麦73(36)9-1可能是一个自发产生的易位系。     

Identification of a 1B/1R wheat-rye chromosome translocation

Summary The common wheat selection 79-4045 was identified as a wheat-rye 1B/1R chromosome translocation line, by means of C-banding patterns and test cross with Chinese Spring double-ditelosomic line. The translocation chromosome consisted of the long arm of wheat chromosome 1B, including its centromere, and the short arm of rye chromosome 1R or tis portion.     

Evaluation of Modified R-B System for Identification of Members of the Family Enterobacteriaceae

In a paired, double-blind study, the modified ("Beckford tube") R-B system was compared with conventional bacteriological procedures for the identification of members of the family Enterobacteriaceae from clinical isolates and stock cultures. The tests in the R-B system yielding positive reactions comparable to those predicted by Ewing's taxonomic classification of Enterobacteriaceae were production of hydrogen sulfide and presence of lysine and ornithine decarboxylasè activities. The test reactions in the R-B system found to be comparable to those in the conventional method were fermentation of glucose, hydrogen sulfide production, and lysine and ornithine decarboxylase activities. The production of gas from glucose was positive in the R-B system more often than in the conventional method; however, the motility test and the production of indole were positive less often in the R-B system. Adequate preliminary identification of the Enterobacteriaceae with the R-B system is enhanced if Simmons' citrate and Christensen's urea tests are used concomitantly. These findings emphasize the manufacturer's instructions that, in interpretation of results, colonial morphology and biochemical reactions must be used concurrently to make an accurate identification.     

Evaluation of the Enterotube System for Identification of Members of the Family Enterobacteriaceae

The Enterotube system was evaluated, in parallel with conventional bacteriological procedures for the identification of members of the family Enterobacteriaceae, by using bacterial strains from a variety of clinical specimens and from stock cultures. Excellent agreement between the two test systems was obtained with the following reactions: hydrogen sulfide, indole, Simmons' citrate, glucose, and lactose. Agreement was not as good (<85%) with the urea, phenylalanine deaminase, and dulcitol reactions. The Enterotube lysine decarboxylase test was unsatisfactory. The Enterotube method will correctly identify strains of the family Enterobacteriaceae approximately 50% of the time; if identification only as Klebsiella-Enterobacter-Serratia group is needed, the method will be correct 85% of the time. On the basis of this evaluation, the Enterotube system appears to be both simple and rapid for the presumptive identification of these bacteria. Because of the limited usefulness of the lysine decarboxylase test, the results obtained by this test system are less reliable than those obtained by conventional methods.     

Efficiency of a Multitest System (Enterotube) for Rapid Identification of Enterobacteriaceae

Enterotube, a multiple-test system which combines nine biochemical tests useful in the identification of members of the family Enterobacteriaceae, was tested and compared with the PathoTec test system in the identification of gram-negative bacilli isolated from clinical specimens. It was found that both the Enterotube and the PathoTec systems rapidly and accurately defined the position of the organisms in the major groups of the family Enterobacteriaceae. Discrepancies were noted between the results of citrate tests on Simmons' citrate-agar and in the Enterotube, as well as between Simmons' citrate-agar and the PathoTec citrate test. It was concluded that the Enterotube system provides a simple, reliable, and rapid method for the presumptive identification of Enterobacteriaceae. The major advantage of the Enterotube is that all tests are done simultaneously by inoculation from a single isolated colony.     

Identification of a 4A/7R and a 7B/4R wheat-rye chromosome translocation

Summary By producing chromosome substitutions with Imperial rye chromosomes 4R (C) and 7R (D) in the wheat cultivar Chinese Spring two spontaneous translocation lines were obtained. One involves segments of wheat chromosome 4A and rye chromosome 7R, the other involves portions of wheat chromosome 7B and rye chromosome 4R     

Numerical Diagnostic Key for the Identification of Enterobacteriaceae

A numerical diagnostic key for enteric organisms is described which permits the identification of typical strains and of biochemical variants with high accuracy. Unknown strains are inoculated into a basic set of five media which permit the testing of eight biochemical reactions. The positive reactions are assigned points, and the score of a strain is added up, after which the identification of the strain is obtained from a table. In many instances, the final identification is obtained with this set of biochemical tests; and, in other instances, a small number of additional tests are required to distinguish between organisms giving the same score in the basic set of biochemical tests. The key permits an accurate, rapid, and economical differentiation of the typical and the more common atypical biotypes of enteric organisms in the clinical laboratory.     

Identification of Members of the Family Enterobacteriaceae by the R-B System

The R-B system was evaluated in parallel with conventional bacteriological procedures for the identification of members of the family Enterobacteriaceae by using bacterial strains from a variety of clinical specimens and from stock cultures. The R-B tests found to be reliable were, in decreasing order, the reactions for phenylalanine deaminase, hydrogen sulfide, and indole, the production of gas from glucose, and the decarboxylation of ornithine. The reactions in the R-B system found to be unreliable were motility, the decarboxylation of lysine, and the fermentation of both glucose and lactose. In addition, the reactions of the R-B system were more difficult to read and interpret than those of the conventional system. On the basis of this evaluation, it was concluded that the R-B system is not an acceptable alternative to the conventional methods in the identification of the Enterobacteriaceae.     

Microfermentation Series for Identification of Single Colonies of Enterobacteriaceae

Microfermentation tests for members of the family Enterobacteriaceae were devised by using agar solutions in disposable, multi-welled, plastic trays. The tests could be made directly from isolated colonies picked from agar plates and represented a considerable saving in time, labor, and materials over the conventional methods. Tests were formulated for determining carbohydrate fermentations, citrate utilization, motility, amino acid decarboxylation, and production of H(2)S, indole, urease, and acetyl-methyl-carbinol.     

Rapid Ornithine Decarboxylase Test for the Identification of Enterobacteriaceae

Conventional methods for detecting ornithine decarboxylase activity require an extended period of incubation. However, with a few simple modifications, accurate results were obtained within a few hours rather than several days. The broth medium was modified, primarily by omitting glucose and by decreasing the pH to 5.5. A 1-ml amount of this broth was inoculated with one colony and then overlaid with sterile mineral oil. Within 2 to 4 hr, the pH increased if ornithine was decarboxylated, thus changing the color of the internal pH indicator to a dark purple. If the amino acid was not decarboxylated, the pH decreased to pH 5.0 to 5.2, enough to give a definite yellow color. With 347 selected clinical isolates, the rapid test gave results identical to those obtained in 1 to 4 days with Moeller's decarboxylase medium. Less reliable results were obtained with Difco's decarboxylase medium with 0.3% agar which was stabinoculated and read after 18 to 24 hr without a mineral oil seal. The rapid ornithine decarboxylase test represents a simple, accurate technique which is well suited for the clinical microbiology laboratory.     

A Propionate-Inducible Expression System for Enteric Bacteria

A series of new expression vectors (pPro) have been constructed for the regulated expression of genes in Escherichia coli. The pPro vectors contain the prpBCDE promoter (P_prpB) responsible for expression of the propionate catabolic genes (prpBCDE) and prpR encoding the positive regulator of this promoter. The efficiency and regulatory properties of the prpR-P_prpB system were measured by placing the gene encoding the green fluorescent protein (gfp) under the control of the inducible P_prpB of E. coli. This system provides homogenous expression in individual cells, highly regulatable expression over a wide range of propionate concentrations, and strong expression (maximal 1,500-fold induction) at high propionate concentrations. Since the prpBCDE promoter has CAP-dependent activation, the prpR-P_prpB system exhibited negligible basal expression by addition of glucose to the medium.     

Identification of Enterobacteriaceae with the Minitek system

A total of 417 strains (361 Enterobacteriaceae, 56 Vibrionaceae) was examined in all the available Minitek system tests. The results were processed through four successive identification schemes devised by the manufacturer and the proportion of strains correctly identified, not identified or incorrectly identified determined for each scheme. From the results, a probability matrix was constructed incorporating all 35 Minitek tests. Test results for each strain were then processed through this matrix to determine its success in identification. From the matrix the order of separating value of the tests was determined. Forty-three of the strains were each tested three times to assess the level of test reproducibility; the corrected error rate was 0.85%.