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PubMed1992

Evaluation of Mast-ID 15 system for identification of fresh clinical isolates of Enterobacteriaceae and Acinetobacter.

AIMS: To assess the accuracy of the Mast-ID 15 system compared with API 20 E for the identification of stock and fresh clinical strains of Enterobacteriaceae and Acinetobacter spp; to compare the accuracy of 19 pin and 36 pin multipoint inoculator heads. METHODS: One hundred frozen stock cultures of Enterobacteriaceae and Acinetobacter spp which had previously been identified by the API 20E were classified by the Mast-ID using 19 and 36 pin multipoint inoculator heads. Reproducibility was determined by testing 36 randomly selected organisms in duplicate. Four hundred and sixty nine consecutive fresh clinical isolates of Enterobacteriaceae and Acinetobacter spp were identified by the Mast-ID using a 36 pin multipoint inoculator and by the API 20E. Reproducibility for the fresh isolates was determined by testing 96 randomly selected strains in duplicate. RESULTS: The Mast-ID 15 identified 82% and 85% of frozen strains to species level and reproducibility was 80% and 86% using 19 and 36 pin inoculator heads, respectively. Of the 469 fresh clinical isolates, the Mast-ID identified 70% of strains to species level; 19% were not identified and 11% were identified incorrectly by comparison with the API 20E. The Mast-ID achieved a reproducibility level of 80% with the fresh clinical isolates. CONCLUSIONS: The use of a 36 pin multipoint inoculator head in preference to the standard 19 pin head for the Mast-ID was advantageous as it allowed greater numbers of strains to be identified at a reduced cost. Unfortunately, in our hands, the Mast-ID system was insufficiently accurate for routine use in the clinical laboratory. Modifications to some of the problematic tests may result in a sufficient increase in accuracy and reproducibility to make the system beneficial in the routine clinical laboratory.

Acinetobacter↗

PubMed1994

Demonstration of the indolepyruvate decarboxylase gene homologue in different auxin-producing species of the Enterobacteriaceae.

Different Enterobacteriaceae were assayed for their ability to produce the plant hormone indole-3-acetate with the aim to study the distribution of the indole-3-pyruvate pathway, which is known to be involved in the production of indole-3-acetate in a root-associated Enterobacter cloacae strain. Other E. cloacae strains, and also Enterobacter agglomerans strains, Pantoea agglomerans, Klebsiella aerogenes, and Klebsiella oxytoca were found to convert tryptophan into indole-3-acetate. As it was also intended to identify the conserved regions of the indole-3-pyruvate decarboxylase, which is involved in producing indole-3-acetate in the E. cloacae strain, oligonucleotide primers were synthesized for different regions of the corresponding gene. One pair of these primers allowed us to amplify a segment of the predicted size by the polymerase chain reaction with DNA of the seven different Enterobacteriaceae that produce indole-3-acetate. Segments of five strains were cloned and sequenced. All sequences showed significant homology to the indole-3-pyruvate decarboxylase gene. As in addition a positive DNA-DNA hybridization signal was detected in the seven strains using the E. cloacae or E. agglomerans segments as a probe, indole-3-acetate biosynthesis is suggested to be catalyzed via the indole-3-pyruvate pathway not only in E. cloacae but also in the other soil-living Enterobacteriaceae. Conserved regions were detected in the indole-3-decarboxylase by alignment of the now-available five different partial sequences. These regions should enable identification of the gene in other bacterial families or even in plants.

Amino Acid Sequence↗

PubMed1996

Distribution of diaminopropane and acetylspermidine in Enterobacteriaceae.

Polyamines of 97 strains (60 species) belonging to 18 genera of the family Enterobacteriaceae were determined by high performance liquid chromatographic analysis. In addition to putrescine and cadavarine, diaminopropane was widely distributed in Enterobacteriaceae and almost ubiquitously within Enterobacter, Pantoea, Erwinia, Leminorella, Proteus, Leclercia, Morganella, Klebsiella, Hafnia, Rahnella, Serratia, and Tatumella species and sporadically within Citrobacter, Escherichia, Moellerella, Providencia, Yokenella, and Yersinia species. Histamine was detected in some cultures of Proteus and Morganella. Agmatine was sporadically spread. Heterogeneity in the occurrence of spermidine was observed within the spermidine-containing cultures. Distribution profiles of 18 genera. Acetylated spermidine was found concomitantly in the spermidine-containing cultures. Distribution profiles of diaminopropane, spermidine, and acetylspermidine in Enterobacteriaceae can serve as a chemotaxonomic marker to distinguish this family from other taxa of the gamma subclass of the class Proteobacteria.

Agmatine↗

PubMed2004

Identification of Enterobacteriaceae from washed and unwashed commercial shell eggs.

To evaluate the effect of processing on the safety and quality of retail shell eggs, a storage study was conducted with unwashed and commercially washed eggs. This work demonstrated that commercial processing decreased microbial contamination of eggshells. To know which species persisted during storage on washed or unwashed eggs, Enterobacteriaceae isolates were selected and identified biochemically. For each of three replications, shell eggs were purchased from a commercial processing plant, transported back to the laboratory, and stored at 4 degrees C. Once a week for 6 weeks, 12 eggs for each treatment (washed and unwashed control) were rinsed in sterile phosphate-buffered saline. A 1-ml aliquot of each sample was plated onto violet red bile glucose agar with overlay and incubated at 37 degrees C for 24 h. Following incubation, plates were observed for colonies characteristic of the family Enterobacteriaceae. A maximum of 10 isolates per positive sample were streaked for isolation before being identified to the genus or species level using commercially available biochemical strips. Although most of the isolates from the unwashed control eggs belonged to the genera Escherichia or Enterobacter, many other genera and species were identified. These included Citrobacter, Klebsiella, Kluyvera, Pantoea, Providencia, Rahnella, Salmonella, Serratia, and Yersinia. Non-Enterobacteriaceae also recovered from the unwashed egg samples included Xanthomonas and Flavimonas. Very few washed egg samples were contaminated with any of these bacteria. These data provide useful information on the effectiveness of processing in removing microorganisms from commercial shell eggs.

Animals↗

PubMed2004

Volatile compounds produced in cheese by Enterobacteriaceae strains of dairy origin.

The formation of volatile compounds in fresh cheese by 10 Enterobacteriaceae strains of dairy origin (4 Hafnia alvei, 2 Serratia liquefaciens, 1 Enterobacter cloacae, 1 Enterobacter sakazakii, and 2 Escherichia coli strains) was investigated. Small cheeses were made from pasteurized cow's milk separately inoculated with 1-3 x 10(3) CFU/ml of each of the Enterobacteriaceae strains, with glucono-8-lactone added to achieve a pH value of 5.2 in the curds. All strains reached counts close to 10(8) CFU/g in 1-day-old cheeses and survived well from day 1 to day 8. Cheeses were analyzed for volatile compounds by gas chromatography-mass spectroscopy, after extraction by dynamic headspace using a purge and trap apparatus. Sixty-one volatile compounds were determined in cheeses, 31 of which were further investigated. Significant increases of aldehydes, sulfur compounds, and aromatic compounds were recorded from 2-h curd to 1-day-old cheese, and of ketones, alcohols, and acids from 2-h curd to 8-day-old cheese. Acetaldehyde, 2-methyl propanal, and 3-methyl butanal predominated among aldehydes; 2,3-butanedione, 2,3-pentanedione, and 3-hydroxy 2-butanone among ketones; ethanol, 2-methyl propanol, and 3-methyl butanol among alcohols; and ethyl acetate among esters. Hierarchical cluster analysis of strains using the data of 31 volatile compounds separated clearly the strain of E. sakazakii, which produced high amounts of volatile compounds, from the other Enterobacteriaceae strains.

Aldehydes↗

PubMed2004

Escherichia coli O157 prevalence and enumeration of aerobic bacteria, Enterobacteriaceae, and Escherichia coli O157 at various steps in commercial beef processing plants.

The effectiveness of current antimicrobial interventions used in reducing the prevalence or load of Escherichia coli O157 and indicator organisms on cattle hides and carcasses at two commercial beef processing plants was evaluated. Sponge sampling of beef cattle was performed at five locations from the initial entry of the animals to the slaughter floor to the exit of carcasses from the "hotbox" cooler. For each sample, E. coli O157 prevalence was determined and total aerobic bacteria, Enterobacteriaceae, and E. coli O157 were enumerated. E. coli O157 was found on 76% of animal hides coming into the plants, but no carcasses leaving the cooler were identified as contaminated with E. coli O157. A positive relationship was seen between the incidence of E. coli O157 in hide samples and that in preevisceration samples. Aerobic plate counts and Enterobacteriaceae counts averaged 7.8 and 6.2 log CFU/100 cm2, respectively, on hides, and 1.4 and 0.4 log CFU/100 cm2, respectively, on chilled carcasses. Aerobic plate counts and Enterobacteriaceae counts on preevisceration carcasses were significantly related to the respective levels on the corresponding hides; the carcasses of animals whose hides carried higher numbers of bacteria were more likely to carry higher numbers of bacteria. Implementation of the sampling protocol described here would allow processors to evaluate the efficacy of on-line antimicrobial interventions and allow industrywide benchmarking of hygienic practices.

Animals↗

PubMed2005

Shell rinse and shell crush methods for the recovery of aerobic microorganisms and enterobacteriaceae from shell eggs.

Recovery of bacteria from shell eggs is important for evaluating the efficacy of processing and the quality and safety of the final product. Shell rinse (SR) techniques are easy to perform and widely used. An alternative sampling method involves crushing and rubbing the shell (CR). To determine the most appropriate method for recovering microorganisms from shell eggs, 358 shell eggs were collected from a commercial egg processor and sampled by SR and CR techniques. Total aerobic mesophiles and Enterobacteriaceae were enumerated on plate count and violet red bile glucose agar plates, respectively. Unwashed, in process, and postprocess eggs were evaluated in the study. Aerobic microorganism prevalence for eggshells sampled was similar for both methods (approximately 100%), but the log CFU per milliliter values were higher in the SR than the CR samples (3.2 and 2.2, respectively). Average Enterobacteriaceae recovery was similar for both methods (45 versus 40% for the SR and CR methods, respectively) when all eggs were considered together. This population was detected more often by SR when unwashed eggs were sampled (90 versus 56% for the SR and CR methods, respectively), equally by SR and CR for in-process eggs (30 versus 29.3% for the SR and CR methods, respectively), but more often by CR for postprocess eggs (10 versus 36% for the SR and CR methods, respectively). The SR technique was easier to perform and recovered larger numbers of aerobic organisms, particularly for unwashed eggs. However, the CR technique was more efficient for recovery of Enterobacteriaceae from postprocess eggs. Stage of shell egg processing may be an important consideration when choosing egg sampling methods.

Animals↗

PubMed2006

Treatments using hot water instead of lactic acid reduce levels of aerobic bacteria and Enterobacteriaceae and reduce the prevalence of Escherichia coil O157:H7 on preevisceration beef carcasses.

Lactic acid has become the most commonly used organic acid for treatment of postevisceration beef carcasses. Many processors have also implemented 2% lactic acid washes on preevisceration carcasses. We previously demonstrated that hot water washing and steam vacuuming are effective carcass interventions. Because of the effectiveness of hot water, we compared its use with that of lactic acid as a preevisceration wash in a commercial setting. A commercial hot water carcass wash cabinet applying 74 degrees C (165 degrees F) water for 5.5 s reduced both aerobic plate counts and Enterobacteriaceae counts by 2.7 log CFU/100 cm2 on preevisceration carcasses. A commercial lactic acid spray cabinet that applied 2% L-lactic acid at approximately 42 degrees C (105 to 110 degrees F) to preevisceration carcasses reduced aerobic plate counts by 1.6 log CFU/100 cm2 and Enterobacteriaceae counts by 1.0 log CFU/100 cm2. When the two cabinets were in use sequentially, i.e., hot water followed by lactic acid, aerobic plate counts were reduced by 2.2 log CFU/100 cm2 and Enterobacteriaceae counts were reduced by 2.5 log CFU/100 cm2. Hot water treatments reduced Escherichia coli O157:H7 prevalence by 81%, and lactic acid treatments reduced E. coli O157:H7 prevalence by 35%, but the two treatments in combination produced a 79% reduction in E. coli O157:H7, a result that was no better than that achieved with hot water alone. These results suggest that hot water would be more beneficial than lactic acid for decontamination of preevisceration beef carcasses.

Animals↗

PubMed2001

Phyloproteomics: species identification of Enterobacteriaceae using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

To evaluate matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) as a tool for rapid identification of common clinical bacterial isolates, we analyzed 25 carefully selected isolates of pathogenic Escherichia coli (E. coli) and additional Enterobacteriaceae members. Organisms were prepared according to clinical microbiological protocols and analyzed with minimal additional processing. Spectra were reproducible from preparation to preparation and comprised 40-100 peaks primarily representing intracellular proteins with masses up to 25 kDa. Spectra of 14 genetically diverse bacteremic isolates of E. coli were compared with isolates representing other genera within the Enterobacteriaceae family. Using a new spectrum comparison algorithm, E. coli isolates were closely related to each other and were readily distinguishable from other Enterobacteriaceae, including Salmonella and Shigella. Presently, the methodology permits the analysis of 40 unknown isolates per hour per instrument. These results suggest that MALDI-ToF MS offers a rapid and reliable approach for performing phyloproteomics i.e., identification of unknown bacterial isolates based on similarities within protein biomarker databases.

Bacterial Proteins↗

PubMed1975

[Nutrition and taxonomy of "enterobacteriaceae" and related bacteria. II. General results and classification (author's transl)].

A study of 186 strains belonging to eleven genera of the family Enterobacteriaceae and to three genera of the family Vibrionaceae has been carried out in order to determine their "versatility" towards 146 organic substrates tested as sole source of carbon and energy. Glucose was the only substrate used by all the strains; gluconate and glycerol were used by respectively 184 and 185 strains; 55 substrates were used by no one strains. The 90 substrates which were used by a fraction of the strains have served to establish a numerical classification of our strains, exclusively relied on these nutritional characters. In using the distinctness coefficient, it was possible to cut the dendrogram into 32 "classes" which may be clustered into 7 "groups" or 2 "sets": the obtained taxa are approximatively similar to the ones which are actually described on the basis of morphological and biochemical characters. The studied strains use 32.7 +/- 11.1 substrates on an average: the "eutrophic Klebsielleae group", which is the most versatile, uses 49.5 substrates on an average, and the "Shigella group", the lease versatile one, only uses 16.5 substrates on an average. The strains of the set 1 (100 strains) use 39.0 substrates on an average: all they belong to the family Enterobacteriaceae. The strains of the set 2 (86 strains) use 25.8 substrates on an average: they belong either to the family of Vibrionaceae, either to any little versatile genus of Enterobacteriaceae, namely Shigella, Proteus and Edwardsiella. The taxonomic inferences of this classification, exclusively established by means of the nutritional characters, are discussed.

Alcohols↗

PubMed2004

Incidence of extended-spectrum beta-lactamases in clinical isolates of the family Enterobacteriaceae in a pediatric hospital.

The incidence of extended-spectrum beta-lactamases (ESBLs) was analyzed in Enterobacteriaceae population circulating in the Upper Silesian Child and Mother Health Center in Katowice (USC&MHC). Altogether 1164 clinical specimens, collected from children hospitalized in 8 different hospital units of USC&MHC were investigated. Five hundred and eighty-five clinical isolates of the family Enterobacteriaceae were identified in specimens collected from 403 patients. Two hundred and twenty-nine Enterobacteriaceae strains (39%) isolated from 162 patients were found to be putative ESBL producers as revealed by double-disc synergy (DDS) test. ESBL activity was the most prevalent in the population of Klebsiella pneumoniae (77%), followed by Klebsiella oxytoca (50%), Serratia marcescens (43%), Escherichia coli (30%), Enterobacter spp. (18%) and Proteus mirabilis (12%). ESBL producers demonstrated also wide resistance to the non-beta-lactam antimicrobial co-trimoxazole (93%) and the aminoglycosides netilmicin (88%), gentamicin (84%) and amikacin (79%).

Child↗

PubMed2006

Simple scheme for identification of common species of enterobacteriaceae.

We propose a simple scheme for the identification of enterobacteriaceae species which routinely necessitates numerous biochemical tests and prolonged time span. In the scheme, family enterobacteriaceae is initially divided into four major groups depending on two important biochemical reactions viz. Lactose fermentation (L) and Methyl red test (MR). Each of the four groups, Group I (L + MR+), Group II (L + MR-), Group III (L- MR-), Group IV (L- MR+) can further be differentiated by using few tests. Eleven genera and 23 species can be identified by this scheme using limited biochemical tests. As many as 990 strains of enterobacteriaceae were subjected to standard biochemical tests and proposed simple scheme for identification. The discrepancy was observed only with 8 atypical strains of E. coli.

Azo Compounds↗

PubMed1991

[Decreasing oxytetracycline resistance in Enterobacteriaceae in veterinary and health conditions after exchange of this substance in animal nutrition].

The national strategy for the use of chemotherapeutics under which there is a distinction between therapeutics and growth promoters for all uses (human medicine, veterinary medicine, animal nutrition) has resulted in a distinct reduction of the selection pressure and a decrease of the OTC resistance among Enterobacteriaceae in the eastern German Länder (formerly GDR) due to a reduction of OTC feeding after 1981 and the final prohibition in 1983. In the following years Enterobacteriaceae were investigated as infectious pathogens for humans and animals using antibiograms. On the one hand, the OTC resistance of porcine and bovine E. coli strains decreased only by 27% and 17%, respectively, during the 1980 to 1988 period, on the other, the OTC resistance of bovine S. typhimurium dropped by 65%, porcine S. typhimurium by 50%, bovine S. dublin by 50%, porcine S. choleraesuis by 30% to a resistance level of about 13% (1989) demonstrating a reduced pressure of resistance. At the same time, OTC resistance became diminished from 50% and 70%, respectively, to 30% also in Enterobacteriaceae (E. coli, Klebsiella) and enterococci isolated from urine and wound infections of patients in the non-agricultural population although therapeutic habits had remained unchanged. The results shown point to a spreading of resistance plasmids which has to be expected far beyond the field of OTC use, for example after the use of antibacterial growth promoters in livestock farming. Control measures should be introduced.

Animal Feed↗

PubMedDate not supplied

Gram-negative identification card for identification of Salmonella, Escherichia coli, and other Enterobacteriaceae isolated from foods: collaborative study.

Twelve laboratories evaluated the Gram-Negative Identification (GNI) Card to identify members of the Enterobacteriaceae. Eighty-four isolates, previously isolated from foods, were used in the collaborative study; the isolates represented 12 genera within the Enterobacteriaceae group. Each collaborator streaked each isolate on tryptic soy agar plates for purity. In the method, plates are incubated 18-24 h at 35 degrees C. Isolated colonies are then subcultured to tryptic soy agar slants and incubated 18-24 h at 35 degrees C. An emulsion is made from the growth on the slant in 1.8 mL 0.45% sodium chloride solution. The GNI Card is filled and placed in a reader/incubator. Isolates are identified and an identification is printed. The Vitek System correctly identified 96.7% of Salmonella sp., 97.0% of Escherichia coli, and an average of 93.8% of the other enteric genera. The method using the Vitek System and GNI Card has been approved interim official first action by AOAC as a screening method for the presumptive identification of Salmonella sp., E. coli, and other Enterobacteriaceae isolated from foods.

Culture Media↗

PubMed1987

[TTE-RAS (Titertek-Enterobac-Ras), a new micromethod for the semi-automatic identification of Enterobacteriaceae within 5 hours].

Titertek-Enterobac-Ras (TTE-RAS), a new semi-automated system for the identification of the Enterobacteriaceae-within five hours, has been evaluated and compared with conventional methods. This note presents the results upon 655 strains, mainly Enterobacteriaceae TTE-RAS provided correct identification for about 92 p. cent and 97 p. cent of Enterobacteriaceae respectively before and after carrying out supplementary tests according to manufacturer's instruction. TTE-RAS gave 97 p. cent specific results for Salmonella. On the other hand, the system correctly identified Aeromonas hydrophila, but not the Gram negative strict aerobic bacteria.

Bacteriological Techniques↗

PubMed1983

[Effect of multiresistance on the minimal inhibitory concentration of N-formimidoyl thienamycin and 6 comparing substances against P. aeruginosa and 4 Enterobacteriaceae species].

The minimal inhibitory concentrations (MICs) of multiresistant strains of P. aeruginosa and four species of Enterobacteriaceae were compared with those of more sensitive ones in N-f-thienamycin and six other antibiotics. The greatest difference between these two differently resistant bacterial populations was found in cefoperazone and the beta-lactamase-instable piperacillin. The latter was used for comparison purposes. The activity of cefsulodin was distinctly reduced in multiresistant strains of P. aeruginosa. Cefotaxime and lamoxactam displayed slightly elevated MICs in multiresistant strains. In N-f-thienamycin a significant difference between the multiresistant and the more sensitive population could not be verified (p greater than 0.05). In this regard it resembles fosfomycin. beta-lactam antibiotics could be ranked by means of correlation analysis of their MICs. Using piperacillin as a reference antibiotic the following order of succession could be arranged according to correlation coefficients in P. aeruginosa: cefoperazone, cefsulodin, lamoxactam, cefotaxime and N-f-thienamycin. For the Enterobacteriaceae species some antibiotics could not be evaluated in this test. Moreover, it was recognized that there was an obvious correlation between the activities of lamoxactam and cefotaxime in P. aeruginosa (r = 0.81) and E. coli (r = 0.80). A correlation was also observed between those of cefsulodin and cefoperazone in P. aeruginosa (r = 0.75). Lower correlation coefficients were seen in N-f-thienamycin with cefoperazone, piperacillin and cefsulodin in P. aeruginosa and with cefoperazone in Klebsiella spec.. Notwithstanding, strains resistant to cefsulodin and cefoperazone were inhibited by concentrations less than or equal to 8 micrograms/ml of N-f-thienamycin. These results showed that in N-f-thienamycin, cefotaxime and lamoxactam multiresistance has only a minimal influence on the MICs of the species of Enterobacteriaceae examined. For therapeutic considerations concerning multiresistance the small differences recognized among these three antibiotics might be without significance. However, N-f-thienamycin was distinctly more active in multiresistant strains of P. aeruginosa than the other beta-lactam antibiotics tested.

Anti-Bacterial Agents↗

PubMed1982

Comparison of the MIC 2000 enteric media with API 20 E and conventional methods for identification of Enterobacteriaceae.

By using MIC 2000 Enteric Identification Media the identification of Enterobacteriaceae was evaluated. 421 strains of Enterobacteriaceae belonging to 23 taxa were identified by MIC 2000, and the conventional method API 20 E, respectively. The results obtained are almost identical, i.e. 96.2% and 96.7%. Besides, the results showed no statistical difference between the two miniaturized systems (p greater than 0.1). The MIC 2000 Enteric Media are capable of identifying the majority of Enterobacteriaceae with a reasonable degree of reliability. This system appears to be efficient and flexible. It is convenient to determine the minimal inhibitory concentration of the tested strain on the same microplate.

Bacteriological Techniques↗

PubMed1980

[Effect of antibiotic therapy in the mother on the colonization of the newborn by enterobacteriaceae (author's transl)].

Two studies were undertaken to determine the effect of antibiotic therapy in the mother on the enterobacteriaceae of the newborn's fecal flora. In the first, mothers were given tetracycline after delivery. Although the effect on the maternal flora was important there was no noticeable effect on the enterobacteriaceae of the newborn although the organisms were predominantly susceptible. In the second, mothers were given ampicillin before delivery and resistant enterobacteriaceae were predominant in all the newborns. In the controls born to untreated mothers susceptible organism still predominate. Antibiotic treatment of the mother has an important effect on the newborn's flora if the treatment is given before delivery.

Ampicillin↗