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PubMed1976

Enterobacteriaceae in the jejunal microflora prevalence and relationship to biochemical and histological evaluations in healthy Colombian men.

When 23 healthy native Southwestern Colombian men were studied to determine the prevalence of Enterobacteriaceae in the jejunal microflora in a sample of thoroughly evaluated tropical inhabitants who were normal by physical examination, chest x-ray, and medical history, 14 of the 23 proved to be Enterobacteriaceae-positive, with counts of 10(3) to 10(9) per milliliter of jejunal aspirate. Thirteen had Escherichia coli, and the fourteenth had Klebsiella pneumoniae. Four had a second species of Enterobacteriaceae associated with E. coli: three were K. pneumoniae, and one was Proteus morganii. Laboratory studies routinely done on all subjects disclosed a total of 18 low biochemical values, 17 of which were associated with 12 of the 14 Enterobacteriaceae-positive subjects: six were low in serum cholesterol, four low in serum vitamin B12, four low in D-xylose excretion, and three low in creatinine coefficient; whereas, the Enterobacteriaceae-negative subjects had normal values for all biochemical tests except for serum vitamin B12 in one case. Nitrogen balance means were significantly different for the two groups: 3.39 g for the Enterobacteriaceae-positive subjects and 1.94 g for the Enterobacteriaceae-negative. No relationship was evident when the histology of the jejunal biopsies was compared with the microbiological or laboratory findings. When the 23 subjects were grouped into those (N = 19) with significant microbial recoveries of any type and those (N = 4) without, the data yielded no meaningful relationships.

Adolescent↗
PubMed2026

Hypervirulence-associated pseudo-compound transposons as fundamental mobile units driving cross-species virulence dissemination in Enterobacteriaceae.

BACKGROUND: The rapid global spread of hypervirulence in Enterobacteriaceae, particularly in carbapenem-resistant Klebsiella pneumoniae, poses a significant public health threat. However, the key genetic vehicles and mechanisms driving horizontal transfer of hypervirulence-associated genes (iucA, iroB, rmpA, rmpA2, and peg-344) remain poorly defined, limiting effective surveillance. METHODS: We performed a large-scale genomic survey of 2,869 virulence-associated plasmid sequences and 2,337 complete Enterobacteriaceae chromosomes. Using comparative genomics and evolutionary analyses, we systematically identified and characterized Hypervirulence-associated Pseudo-Compound Transposons (Hva-PCTs), defined as structured mobile elements in which hypervirulence-associated genes are flanked by insertion sequences. RESULTS: Our results demonstrate that hypervirulence-associated genes are transmitted primarily as discrete IS-bounded units, which we term Hva-PCTs. We identified 29 distinct plasmid-borne Hva-PCTs (pHva-PCTs) and 30 chromosomal Hva-PCTs (cHva-PCTs). These modules show clear species-specific patterns: iucA/iroB-associated Hva-PCTs mainly originate in Escherichia coli and spread through IncFIB-containing multi-replicon plasmids (commonly combined with IncFIC(FII) and/or IncFII, while rmpA/rmpA2/peg-344-containing modules originate in K. pneumoniae and are disseminated via IncHI1B/repB plasmids. Three Hva-PCTs were detected on both plasmids and chromosomes (xHva-PCTs). In one clinical K. pneumoniae isolate (LS356), the identical composite module was present on both replicons. Simpler sub-modules, such as ISKqu3-rmpA2-iucA_1-IS102 and IS102-rmpA-peg-344-iroB_1-IS1A, frequently co-occur on the same plasmid; when positioned in tandem, they reconstitute the full composite structure. This assembly pattern is further supported by a partial duplication event in plasmid pP901. CD-HIT clustering (80% nucleotide identity and 90% coverage) showed that 13 of 22 major clusters contained both plasmid and chromosomal copies, with intra-cluster identities >80% across multiple sequence types and host species. CONCLUSION: Hypervirulence-associated genes in Enterobacteriaceae are disseminated mainly as IS-flanked Hva-PCTs rather than solely through intact virulence plasmids. These modules exhibit strong but not absolute host specificity. The presence of identical Hva-PCTs on plasmids and chromosomes suggests inter-replicon mobility, while their stepwise assembly from simpler sub-modules highlights modular accretion as a key evolutionary process. Tracking Hva-PCTs as distinct mobile units may complement existing plasmid- and gene-centric surveillance approaches for hypervirulent and convergent strains. Experimental validation of their transposition activity and phenotypic effects is still required.

Virulence↗
PubMed1979

[An oral enteritis-vaccine composed of twelve heat inactivated Enterobacteriaceae. 1. Communication: Theoretical and epidemiological considerations (author's transl)].

The infectious diseases of the human intestinal tract which are caused by bacteria must be distinguished into two groups on account of their different pathogenesis: the cyclic infections (typhoid fever, parathyphoid fever) and the local infections (cholera, dysentery, Salmonella enteritis, dyspepsia coli infections). The local infections of the intestine do not cause a systemic but only a local immunity of the intestinal mucosa. It is necessary therefore to induce local immunity as active immunoprophylaxis by orally administering inactivated antigens. The twelve-fold enteritis vaccine consists of full antigens of 6 Salmonella strains, 2 Shigella strains, and 4 enteropathogenic coli strains pretreated by heat-inactivation (3 min/100 degrees C). The following should be considered as indication to effect active immunoprophylaxis against enteritis: Travelling into tropical and subtropical countries, people in emergency areas, children in developing countries, workers in food industries, secondary hospital infections, and carriers. The active mouse protection test revealed that oral immunization with enterobacteriaceae does not only deliver the well-known specific effect but also a non-specific effect which included the protection against other related enterobacteriaceae. Moreover, the specific component of the combined vaccine is enhanced by heterologous components. The resulting synergism or the adjuvantal effect, respectively, allows to employ a relatively limited number of germs which are selected on the basis of high pathogenicity, good immunogenicity, and great frequency. The first field trial with the twelve-fold vaccine was completed successfully: Following an infection with Salmonella which affected the employees of a fowl slaughtery, eight different species could be demonstrated; the above described polyvalent vaccine was orally administered and proved to be successful. The latter case clearly demonstrates the fast-acting effect of the vaccine on account of the heterologous bacterial antigens contained therein. 51 out of 60 Salmonella carriers excreted germs of a different antigen pattern not contained in the vaccine. However, the good results obtained showed that the species chosen for the vaccine were still sufficiently effective to cover the wide spectrum of other species of related enterobacteriaceae.

Administration, Oral↗
PubMed1977

The use of bile - esculin agar for the taxonomic classification of the family Enterobacteriaceae.

Bile-esculin medium has been used for many years for the presumptive identification of group D Streptococcus. The test is based on the ability of a bacterium to grow in the presence of 40% bile and produce esculinase. 2935 strains of Enterobacteriaceae were inoculated onto bile-esculin agar slants and incubated at 35 C. Esculin hydrolysis was determined after 24 and 48 hours. At 24 hours of incubation esculin hydrolysis was limited to the genera Klebsiella, Enterobacter, Serratia, and the species P. vulgaris, P. rettgeri, and C. diversus. Not all strains of these species were positive, however. All other members of the family were negative. At 48 hours of incubation 37% of E. coli gave a positive reaction; all other Enterobacteriaceae which were negative at 24 hours remained negative. Esculin hydrolysis is a valuable test for the taxonomic classification of the family Enterobacteriaceae.

Bacteriological Techniques↗
PubMed1977

The reliability of the examination of foods, processed for safety, for enteric pathogens and Enterobacteriaceae: a mathematical and ecological study.

Because of the paucity of quantitative data on numbers of other enteric pathogens in food, the reliability of the examination of processed foods for Enterobacteriaceae was estimated taking Salmonella as a model. For this purpose an assessment was carried out of the risk of accepting Salmonella contaminated consignments of foods, despite a negative outcome of (i) examination of 1.5 kg samples for Salmonella; (ii) examination of one or two 1 g samples for Enterobacteriaceae; (iii) simultaneous application of both tests. The computations were based on the results of the examination of 6830 samples of dried foods, processed for safety, out of a total of 18170 samples.Only 69 samples permitted the exact calculation of the epsilon-factor, defined as c.f.u./g of Enterobacteriaceae/c.f.u./g of Salmonella; 4642 were positive for the former group but ;free' from Salmonella, and the rest were negative in both tests. Numbers of c.f.u./g for both groups, and hence the epsilon-factors, varied widely between commodities and also between different consignments of the same food product. The average for epsilon amounted to 5.8 x 10(3), far from the base-line value of 0.75 x 10(3) assessed earlier. In only 0.1% of samples did the Enterobacteriaceae test fail to achieve the required consumer protection.This investigation therefore substantiates that testing foods processed for safety by examining accurately chosen quantities for ecologically well selected and taxonomically thoroughly defined index organisms is a most effective procedure in terms both of consumer protection and simplicity of examination without compelling the food industry to achieve hardly attainable microbiological quality standards.

Bacteriological Techniques↗
PubMed1979

Antigenic cross-reactivity of major outer membrane proteins in enterobacteriaceae species.

The protein constituents in the outer membrane (OM) of several serotypes of Escherichia coli and some other Enterobacteriaceae cross-reacted antigenically. Solubilized OM preparations of these bacteria were applied in interfacial precipitin tests to antisera elicited in rabbits against whole bacterial cells, absorbed with their appropriate lipopolysaccharide before testing. The resulting immunecomplexes were analysed on polyacrylamide gels. Protein profiles of the immunoprecipitates showed a considerable antigenic cross-reactivity of outer membrane proteins between most E. coli serotypes. Cross-reactivity, though substantially lower, was also found with OM from three other Enterobacteriaceae species, but was not detectable with Pseudomonas aeruginosa OM. When OM preparations were solubilized at room temperature, the peptidoglycan-bound proteins in the molecular weight range 37,000 to 41,000 predominated in the protein profiles of the immunecomplexes. In profiles of immunecomplexes obtained with boiled OM preparations, a heat-modifiable protein (mol. wt 33,000) predominated. The major OM proteins of the Gram-negative bacterium may therefore play a role as common surface antigens of the family of Enterobacteriaceae.

Antigen-Antibody Complex↗
PubMed1976

Comparative activity of netilmicin, gentamicin, amikacin, and tobramycin against Pseudomonas aeruginosa and Enterobacteriaceae.

Netilmicin (Sch 20569), a semisynthetic aminoglycoside antibiotic, was compared with gentamicin, tobramycin, and amikacin against 242 clinical isolates of Pseudomonas and Enterobacteriaceae. The minimum inhibitory concentration (MIC) was determined in both solid and liquid media. Netilmicin exhibited typical aminoglycoside properties, such as little effect of inoculum size on MIC, relatively small gap between MIC and minimum bactericidal concentration, and potentiation of anti-Pseudomonas activity in the presence of carbenicillin. Netilmicin provided no advantage in antimicrobial activity over gentamicin for either Pseudomonas or Enterobacteriaceae. Nearly complete cross-resistance to netilmicin was encountered with isolates resistant to gentamicin in either solid or liquid media. Netilmicin was less active than gentamicin against isolates of Pseudomonas and Providencia. Major discrepancies between MIC values determined in agar as opposed to those determined in broth were encountered for most isolates of Pseudomonas but also, depending upon antibiotic tested, for between 15 and 40% of isolates of Enterobacteriaceae. This new aminoglycoside agent will be useful clinically only if it is shown to be significantly less toxic than presently available analogues.

Amikacin↗
PubMed1976

Antibacterial activity of selected beta-lactam and aminoglycoside antibiotics against cephalothin-resistant Enterobacteriaceae.

The in vitro antibacterial activity of four beta-lactam antibiotics (cefatrizine [BL-S640], cefamandole, cefoxitin, and carbenicillin) and three aminoglycosides (amikacin, gentamicin, and tobramycin) was determined against 197 strains of cephalothin-resistant Enterobacteriaceae. Eighty strains were found to be gentamicin-sensitive, and 117 were found to be gentamicin-resistant. Carbenicillin was the most active beta-lactam antibiotic against gentamicin-sensitive Serratia marcescens and Enterobacter spp. Cefoxitin was the most active beta-lactam antibiotic against the remaining gentamicin-sensitive and -resistant Enterobacteriaceae, including Providencia stuartii and indole-positive Proteus spp. Cefatrizine exhibited little activity against the organisms studied. Cefamandole was less active than cefoxitin and carbenicillin. Amikacin was the most effective agent in vitro. With the exception of S. marcescens, cefoxitin appeared to be the next most promising agent in vitro against gentamicin- and cephalothin-resistant Enterobacteriaceae.

Aminoglycosides↗
PubMed1977

Esculin hydrolysis by Enterobacteriaceae.

Literature reports disagree concerning esculin hydrolysis in the family Enterobacteriaceae. A total of 2,490 strains of the family were investigated for esculin hydrolysis by two methods, the esculin spot test and the PathoTec incubation strip, which measures constitutive enzyme, and five growth-supporting methods, which determine both constitutive and inducible enzymes. The five growth-supporting media studied were: Vaughn-Levine, the standard esculin hydrolysis medium (P. R. Edwards and W. H. Ewing, Identification of Enterobacteriaceae, 3rd ed., 1972); Vaughn-Levine without iron; Vaughn-Levine without Andrade's indicator; and bile-esculin medium. Growth media were incubated at 35 degrees C and checked every 24 h for 120 h. On growth media, 0.3% of Escherichia coli were positive in 24 h, 34% in 48 h, and 61% in 120 h. No strains were positive on the "nongrowth" tests. It appeared that the esculin hydrolysis enzyme(s) of E. coli was inducible rather than constitutive. All esculin hydrolyzers, which yielded positive tests on "constitutive tests" and 24-h tests, were limited to the genera Klebsiella, Enterobacter, and Serratia and species of Proteus vulgaris, Proteus rettgeri, and Citrobacter diversus. When used with standardized inoculum size and incubation time, the esculin hydrolysis test is very useful for differentiation within the family Enterobacteriaceae.

Citrobacter↗
PubMed1976

[Assaying the Enterobacteriaceae contents of fishmeal as criterion for absence of Salmonellae (author's transl)].

1. During comparison of different test systems for examination on salmonellae the routine method revealed only 2 of 35 fishmeal consignments as positive. In second streaks after 48 hours on selective media 5 more consignments were discovered to contain samonellae. Using pre-enrichment culture gave evidence for 11 positive consignments altogether. 2. The pre-enrichment method led to about 20 times higher yields in isolating enterobacteriaceae than the usual enrichment method. 3. Neither salmonellae nor enterobacteriaceae statistically showed uniform distribution in fishmeal. In addition, with the aid of the variance test of homogenity, an extremely heterogeneous ratio of the both germ types to each other was proved. Consequently, assaying the enterobacteriaceae contents is not suitable to draw any reliable conclusions upon the salmonellae contents of fishmeal. 4. Assuming a constant contamination rate of 400.000 salmonella units in consignments of 100 tons and testing them by the required 47 random samples, it can be deduced from probability calculus that only about 1 per cent of a larger number of consignments will fail to be recognized as positive. This risk of acceptance increases progressively with diminishing rate of salmonella contamination. Thus about 80 per cent of consignments containing even as much as 20.000 salmonella units will be accepted as "false negatives". 5. Because of the usually low contamination rates of fishmeal it is an erroneous idea to increase the efficiency of assay by modifying the sample sizes in relation to the shipping weight. For this reason the orders for fishmeal examination in the Federal Republic of Germany should be revised. It is suggested to take at least the number of samples required hitherto for consignments of 100 tons. By this without doubt the risk of accepting "false negatives" can be reduced significantly, but it remains still impossible to recognite every contaminated fishmeal consignment and to stop its acceptance by bacteriological examination. 6. From practical examinations and theoretical considerations it is derivable that one has either to tolerate a low degree of salmonella contamination which hygienically may be without any importance or to treat every imported fishmeal by irradiation or heat pelleting.

Bacteriological Techniques↗
PubMed1979

Clinical evaluation of the MICRO-ID, API 20E, and conventional media systems for identification of Enterobacteriacea.

MICRO-ID (General Diagnostics, Morris Plains, N.J.) is a new kit system designed for the identification of Enterobacteriaceae in 4 h. It consists of 15 biochemical tests of paper disks. Each test is in its own compartment in a molded plastic tray. Only one reagent need be added to the system (2 drops of 20% KOH, which is added to the Voges-Proskauer test). Based on the pattern of positive and negative biochemical test results, a five-digit octal code number is calculated. An identification is derived from a computer-generated identification manual. A study was conducted to compare three systems-the MICRO-ID 4-h and the API 20E (Analytab Products Inc., Plainview, N.Y.) 18- to 24-h systems and a conventional media system-to measure the ability of each to identify members of the family Enterobacteriaceae. Comparison tables, rather than simple percentage agreement tables, were generated to define the particular strengths and weaknesses of each system and allow the laboratory to best use the data. The MICRO-ID compared quite favorably with conventional media. MICRO-ID yielded incorrect identifications with 1.5% of the isolates tested (API 20E, 4.7% misidentification rate). Half the MICRO-ID misidentifications occurred when the system identified a Citrobacter diversus as a lysine-negative Escherichia coli; all gave one octal number. A direct comparison of the MICRO-ID and API 20E was of limited value because percentage agreements were merely the sums of the errors of each. The ease of inoculation, the requirement for the addition of only one reagent, and the 4-h capability make the MICRO-ID system an extremely attractive development in the field of bacterial identification.

Bacteriological Techniques↗
PubMed1979

Evaluation of the modified Micro-ID system for identification of Enterobacteriaceae.

Micro-ID is a system designed to identify the Enterobacteriaceae by utilizing reagent-impregnated disks for 15 biochemical tests. Since its initial evaluations, the system has undergone modification in formulation and in its computer data base. In a dual-center evaluation, 306 isolates of Enterobacteriaceae were tested: 145 common and typical isolates at the Mayo Clinic and 161 unusual or atypical isolates at the Center for Disease Control. Each laboratory also exchanged 50 cultures to test the system's reproducibility. Micro-ID correctly identified 142 (98%) of the common clinical isolates and 123 (76%) of the unusual or atypical organisms. However, in this latter group, three species tested were not in the system's data base. When these organisms were deleted from the analysis, 138 of 146 (95%) of the unusual or atypical isolates were correctly identified. Analysis of the 100 isolates identified in duplicate revealed 93% reproducibility of genus and species identification and 62% reproducibility of octal code numbers. Of the 31 strains with the same identification but different code numbers, 74% differed in only one biochemical test.

Bacteriological Techniques↗
PubMed1979

Silver-resistant Enterobacteriaceae from hospital patients.

The inclusion of agar medium containing 0.5 mM AgNO3 in the hospital laboratory replicating system for routine antibiotic-susceptibility determinations resulted in identification of species of Enterobacteriaceae (Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Proteus mirabilis, and Citrobacter freundii) with silver resistance. Since the study began in October, 1975, 11 in-hospital patients receiving silver sulfadiazine for burn wound prophylaxis have yielded silver-resistant bacteria from their infected burns. During this treatment routine burn-site cultures from these patients yielded 230 isolates of Enterobacteriaceae, including 211 which were sulfonamide-resistant, 97 of which were also silver-resistant, and 38 of which were untested for silver resistance. Seven silver-resistant but sulfonamide-sensitive isolates were incidentally recovered from respiratory specimens from four nonburn patients with silver tracheostomy tubes, one silver-resistant sulfonamide-sensitive isolate was recovered from a small infected burn on the foot of an Emergency Room patient. Previous treatment of this burn was unknown. Representative AgNO3-resistant E. coli isolates from four patients were serologically untypable. Serotyping of representative isolates of K. pneumoniae showed a diversity of types except from two patients who had been in the same ward at the same time.

Burns↗
PubMed1979

Enterobacteriaceae infections in man and animals and detection of their natural reservoirs in India.

The study presents a summary of the results regarding the presence of Enterobacteriaceae in domestic animals and birds, endeavouring to explain the origin of the infections. Wild reptiles, amphibians, mammalia, rodents and birds were examined into the demonstration of Enterobacteriaceae, whereby particular attention was paid to the following species: Arizona, Edwardsiella, Citrobacter, Enterobacter, Serratia, Yersinia and Erwinia.

Animal Population Groups↗
PubMed1978

Iron requirement and chelator production of staphylococci, Streptococcus faecalis and enterobacteriaceae.

The effect of iron deprivation on growth of 101 aerobic strains of gram-positive and gram-negative bacteria was studied on agar media in the presence of various concentrations of the synthetic iron chelator ethylene diamine diorthohydroxyphenyl acetic acid (EDDA) and the iron binding protein transferrin. Growth of Staphylococcus epidermidis was inhibited by 15 mM EDDA and 1.5 mM transferrin. Staphylococcus aureus was only inhibited by 44 mM EDDA and not by transferrin. None of the strains of S. faecalis was inhibited. The majority of the enterobacteriaceae (E. coli, Salmonella spp, Klebsiella spp) was inhibited by 44 mM EDDA and 1.5 mM transferrin. The relation between susceptibility and concentration of EDDA and transferrin was expressed as S-value for each species. Iron supply with various iron compounds could restore the effects of inhibition. In all species except in S. faecalis iron chelator production could be demonstrated, using indicator plates of media containing EDDA and flooded with 10(4)--10(5) colony forming units of indicator organisms. The iron chelator of both S. epidermidis and S. aureus could stimulate growth of S. epidermidis, but not that of enterobacteriaceae. Iron chelators from all gram-negative bacteria were functionally interchangeable, but did not stimulate growth of gram-positive bacteria.

Chelating Agents↗
PubMed1977

Interaction of blood with enterobacteriaceae: hemolysis, hemagglutination, fibrinolysis.

One thousand eighty enterobacteriaceae, isolated during a period of three months, were tested for the presence of hemolysis, hemagglutination and fibrinolysis on human erythrocytes and plasma. The fibrinolysis on human erythrocytes and plasma. The Escherichia coli have the greatest diversity of action on the blood. Of 481 strains, 32.9% were hemolytic, 21.9% fibrinolytic, and 10.5% hemagglutinating. Of 190 strains of Klebsiella, 95.2% were fibrinolytic. Of 53 strains of Enterobacter, 90.6% were fibrinolytic, and of 53 strains of Enterobacter hafniae, 66% were fibrinolytic. Of seven strains of Serratia, 28.5% were hemolytic and 14.3% fibrinolytic. The Proteae were mostly fibrinolytic: 60.5% of 141 strains, with three strains being hemolytic and fibrinolytic. Of 19 strains of Citrobacter, 52.6% were fibrinolytic only. Providenica, Salmonella and Shigella did not interact with human blood. The patterns of blood interactions with the different species of Enterobacteriaceae are of value in the evaluation of the pathogenesis of the infections and septicemias they cause. Their study is an essential step in the understanding of the sequence of events leading to severe disturbances of the blood-clotting mechanism.

Enterobacteriaceae↗
PubMed1979

Selection of small-colony variants of Enterobacteriaceae by in vitro exposure to aminoglycosides: pathogenicity for experimental animals.

Small-colony variants of gram-negative genera of Enterobacteriaceae were selected by in vitro exposure to gentamicin. These variants were shown to have decreased susceptibility in vitro to aminoglycosides. They were lethal for mice following intraperitoneal injection, with the LD50 (50% lethal dose) being the same as, or slightly less than, that for the parent organism. Variants of strain no. 2401 of Proteus mirabilis caused urinary tract infection in mice after implantation into their bladders. Although the variants grew somewhat less rapidly than did parent organisms, both parent and variant colonies alkalinized urine at the same rate. Electron microscopic study showed no differences between colonies of parents and variants. These studies indicated that small-colony variants of Enterobacteriaceae are pathogenic for experimental animals. Further, they may cause disease in humans and should not necessarily be regarded as only laboratory curiosities.

Aminoglycosides↗
PubMed1976

A heterophile system in human renal transplantation. V. Relationship of heterophile transplantation antigen and common antigen of Enterobacteriaceae.

Heterophile transplantation antigen and common antigen of Enterobacteriaceae appear serologically to be separate specificities. However, both antigens are common to Enterobacteriaceae, rat erythrocytes, and some human kidneys. Both antigens are obtained from various tissues by the same chemical procedure. Immunity to each antigen is frequently produced by renal transplantation. We suggest that the antigens are either separate molecules which are similar in chemical structure in the region of the antigenic determinant as well as in tissue distribution or separate reactive sites located on the same molecule. The possibility that common antigen may be a human alloantigen raises theoretical possibilities relative to susceptibility to infection and pyelonephritis, as well as to its relationship to histocompatibility.

Animals↗