Abstract: Disclosed herein are methods for making large amounts of highly pure colonization factors. The methods of the present invention differ from prior art methods in that host cells which express the colonization factor of interest are cultured in media comprising more than about 50 ?g/l of an antibiotic, the media is centrifuged and then filtered with a 0.2 ?m filter tangential flow cartridge and a 300,000 MW cut-off filter, and a divalent cation is added. As disclosed herein the colonization factors made by the method of the present invention may be used in pharmaceutical compositions and methods for treating or preventing enterotoxigenic Escherichia coli infections.

Abstract: The invention relates to a strain suitable for producing a live, orally applicable Escherichia coli vaccine for the prevention of post-weaning diarrhoea in pigs, and the procedure suitable for producing that strain. The essence of the strain is that the enterotoxin-free and originally wild-type Escherichia coli strain simultaneously produces two adhesive fimbriae (F4 and F18), whereas the essence of the procedure is that the enterotoxin-producing ability if the wild, pathogenic, enterotoxigenic Escherichia coli strain originally capable of producing enterotoxins and F18 fimbriae is abolished by a genetic intervention while retaining the ability of the strain to produce F18 fimbria facilitating adhesion to the small intestinal wall of weaned piglets, and subsequently the strain thus modified is rendered capable of producing a further surface adhesion fimbria (F4).

Abstract: The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Streptococcus equisimilis hyaluronate synthase (seHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product. Hyaluronate is also known as hyaluronic acid or hyaluronan.

Abstract: The present invention is related to microbiology and forms part of a system for rapid microbiological diagnosis. The invention allows detection of turbidimetric changes due to microbial growth, using equipment comprised of two main devices: a static turbidimetric minireader and a microflow sensor which is fed by a peristaltic pump; this equipment is coupled to a microcomputer with a program package for acquisition, processing and formation of databases used in generating necessary reports. The diagnostic kit has a glass vial with culture medium and a polymer with derepressive activity and two additional substrates for E.coli identification, as well as a set of antibiotic discs arranged in a strip for antibiogram determination from previously isolated colonies or samples obtained directly from their sources, allowing detection of urinary tract infections from direct samples of urine, and additionally simultaneous identification of E.coli.

Abstract: Disclosed are methods and devices for detection of bacteria based on recognition and infection of one or more selected strains of bacteria with bacteriophage genetically modified to cause production of an inducer molecule in the bacterium following phage infection. The inducer molecule is released from the infected bacterium and is detected by genetically modified bacterial bioreporter cells designed to emit bioluminescence upon stimulation by the inducer. Autoamplification of the bioluminescent signal permits detection of low levels of bacteria without sample enrichment. Also disclosed are methods of detection for select bacteria, and kits for detection of select bacteria based on the described technology.

Abstract: Fungi and bacteria can be detected and rapidly quantified by using the nucleotide sequences taught here that are specific to the particular species or group of species of fungi or bacteria. Use of the sequences can be made with fluorescent labeled probes, such as in the TaqMan™ system which produces real time detection of polymerase chain reaction (PCR) products. Other methods of detection and quantification based on these sequences include hybridization, conventional PCR or other molecular techniques.

Abstract: The present invention concerns methods of testing water for microbe contamination. The methods of the invention comprise supplementing existing methods with assays using specific reagents such as monoclonal antibodies. The invention also concerns a device for use in the methods of the invention.

Abstract: An industrially more advantageous method for the production of metabolites biologically synthesized via phosphoribosyl pyrophosphate (PRPP) is provided, making use of metabolically modified strains in which transketolase activity is deficient or reduced in comparison with the parent strain.

Abstract: An assay for compounds useful in the treatment of a bacterial induced coagulation disorder has the following steps: a) incubating a plasma sample with a strain of bacteria; b) adding a compound to be assayed to the plasma sample before, during or after step (a); c) conducting an activated partial thromboplastin time test; d) determining the clotting time.

Abstract: This invention relates to a method for removing fumarase activity from a microorganism or processed product thereof having ethylenediamine-N,N'-disuccinic acid ethylenediamine lyase activity, which includes treating the microorganism or processed product thereof with an aqueous alkaline solution at a pH of 8.0 to 10.5 in the presence of at least one salt with a concentration of 5 mM to 1000 mM. The salt is preferably selected from the group consisting of sodium, potassium, ammonium and C.sub.2-6 alkanediamine salts of boric acid, phosphoric acid, hydrochloric acid, sulfuric acid, acetic acid, oxalic acid, fumaric acid, maleic acid and ethylenediamine-N,N'-disuccinic acid, and mixtures thereof.

Abstract: A self-contained incubator for growth of microorganism kit and methods for use of such a kit are provided. The kit and methods may detect the presence of microorganisms and may utilie a microorganism growth and indicator medium provided in a sample container along with a heat source, preferably generating heat through chemical means, and optionally heat shields, allowing for on-site testing of a microorganism present in a sample. The sample container may also include a removable vessel cap that includes a barrier separating the sample from a material capable of disinfecting the sample, thereby preventing contact of the sample and the material for a desired time period. The vessel cap may also be used independently in other applications.

Abstract: The present invention is directed to a method of using a liquid matrix containing a black body light absorbing powder to facilitate the analysis of biomarkers from representative microorganisms by laser desorption mass spectrometry. Both an IR laser (1064 nm) and a UV laser (337 nm) were shown to be compatible and both time-of-flight and Fourier-transform mass analyzer were used. In the present implementation gram negative and gram positive bacteria were suspended in a methanol:chloroform solution and added to a cobalt/glycerol matrix, S/N, sensitivity and sampling time are greatly enhanced for polar lipid biomarkers.

Abstract: The present disclosure reports a thin film culture plate device including i) a self-supporting, waterproof substrate containing a layer of a unique reconstitutable culture medium made of nutrients for growing microorganisms, and a mixture of gelling agents which are prepared in granular form by agglomerating the nutrients and mixture of gelling agents in the presence of an aqueous binder and ii) a cover sheet adhered to a portion of the substrate. Methods to make agglomerated medium particles are also reported.

Abstract: A two stage enzymatic method for the detection of coliform bacteria or E. coli wherein bacteria are concentrated on a membrane filter. This filter is placed on a growth medium containing nutrients, including preferably minerals, a protein hydrolysate and a sugar, preferably maltose or a polyalcohol, preferably mannitol, an inducer of a marker enzyme, in particular .beta.-galactosidase or .beta.-glucuronidase and inhibitors of the growth of competing bacteria. After a preincubation step, the filter is placed on an assay medium containing a fluorogenic or chemiluminogenic enzyme substrate and a membrane permeabilizer. The membrane filter and the assay medium are incubated to allow cleavage of the enzyme substrate producing fluorescent or chemiluminescent microcolonies on the membrane filter after triggering of light emission.

Abstract: A selective culture medium which permits simultaneous detection of total coliform and Escherichia coli in a test sample with a single growth phase incubation period. The culture medium includes the required components of: (i) carbon nutrients, (ii) a nitrogen nutrient, (iii) a source of metabolizable potassium, (iv) a source of metabolizable phosphate, (v) vitamins, (vi) minerals, (vii) amino acids, (viii) sodium pyruvate, (ix) a bactericidal system selective for non-coliform bacteria which includes methylene blue, erythromycin and an azide, and (x) a sensible indicator selectively metabolized by Escherichia coli to the exclusion of other coliforms.

Abstract: A test kit and method for the highly sensitive detection of specific analytes in a sample is provided. The presence of the analyte in the sample results in a decrease in the concentration of a growth inhibiting substance leading to proliferation of cells in the region of the analyte. The presence or absence of the analyte is determined by detecting the presence of increased numbers of cells. Assay sensitivity is accounted for by the exponential amplification of cell number that occurs during cell proliferation in the presence of analyte.

Abstract: The invention provides a cosmetic or dermatological method for detecting and/or selectively quantifying individual microorganisms, and/or whole groups of microorganisms, which are present on human or animal skin, comprising the steps ofremoving a sample of the microflora of the human or animal skin,treating the sample with a deinhibiting medium, adding the treated sample to a culture medium which exhibits favorable growth conditions for a defined group of microorganisms but unfavorable growth conditions for other microorganisms, to produce a selective culture, and incubating the selective culture over a sufficiently long period of time, to allow only the group of microorganisms for which the culture medium exhibits favorable growth conditions the opportunity to multiply, in association with metabolic products, in particular CO.sub.

Abstract: A process for producing L-leucine, which includes incubating an L-leucine-productive microorganism belonging to the genus Corynebacterium, Escherichia, Brevibacterium, or Microbacterium in a culture medium and reacting the resulting cells with saccharides and acetic acid or its salt to form and accumulate L-leucine in the reaction solution. The process improves the amount of L-leucine accumulated and decreases formation of amino acid byproducts.

Abstract: A method for determining the sensitivity of at least one nonparaffinophilic microorganism from a specimen obtained from a patient to an antimicrobial agent. The method includes providing at least one receptacle containing an aqueous solution that does not contain a carbon source and inoculating the solution with the specimen. The method further includes placing into the receptacle (i) a slide having bound thereto a carbon source and (ii) a predetermined quantity of an antimicrobial agent to be tested. By observing the nonparaffinophilic microorganism growth or lack thereof on the slide, it can be determined whether the predetermined quantity of the antimicrobial agent is effective in inhibiting growth of the nonparaffinophilic microorganism on the slide. An associated apparatus is also disclosed.

Abstract: A selective culture medium which permits simultaneous detection of total coliform and Escherichia coli in a test sample with a single growth phase incubation period.

Abstract: This invention generally relates to products and processes used to determine the presence of Enterobacteriaceae in a sample and particularly relates to a bacterial culture medium which may be used in products and processes to allow early detection and enumeration of Enterobacteriaceae in a sample. The bacterial culture medium which facilitates the early detection and enumeration of Enterobacteriaceae contains a selected amount of glucose, pH indicator and buffer which prevent diffusion of colored indicator zones associated with growing bacteria in the medium.

Abstract: Oligonucleotides (SEQ ID NOs 1-8) selectively hybridizable with a specific gene of Vibro parahaemolyticus, oligonucleotides (SEQ ID NOs 9-13) selectively hybridizable with the LT gene of toxigenic Escherchia coil, oligonucleotides (SEQ ID NOs 14-21) selectively hybrizable with the STh or STp gene of toxigenic Escherchia coil, oligonucleotides (SEQ ID NOs 22-47) selectively hybridizable with the entA, B, C, or D gene of Staphylococcus aureus, or oligonucleotides (SEQ ID NOs 48-53) selectively hybridizable with the entE gene of Staplyloccus aureus are prepared and used as primers for gene amplification to thereby selectively detect only respective microorganisms causing food poisoning.

Abstract: Oligonucleotides (SEQ ID NOs 1-8) selectively hybridizable with a specific gene of Vibro parahaemolyticus, oligonucleotides (SEQ ID NOs 9-13) selectively hybridizable with the LT gene of toxigenic Escherchia coil, oligonucleotides (SEQ ID NOs 14-21) selectively hybrizable with the STh or STp gene of toxigenic Escherchia coil, oligonucleotides (SEQ ID NOs 22-47) selectively hybridizable with the entA, B, C, or D gene of Staphylococcus aureus, or oligonucleotides (SEQ ID NOs 48-53) selectively hybridizable with the entE gene of Staplyloccus aureus are prepared and used as primers for gene amplification to thereby selectively detect only respective microorganisms causing food poisoning.

Abstract: A test kit and method for the amplification and detection of specific antigen cells using a probe. The method includes reacting the probe-specific cells with enzyme-conjugated molecules to form separate molecules. The specific antigen cells are mixed with a selected antibiotic which antibiotic is adversely affected by the enzyme in the reporter molecules and incubating the mixture to promote a bacterial chain reaction forming satellite colonies of bacteria microcolonies about the specific cells which amplifies the cells. The method then includes detecting the amplified probe-specific cells by observing the satellite colonies.

Abstract: The present invention provides a mutant and discloses a process for producing L-glutamic acid by fermentation using a microorganism belonging to the genus Escherichia. The L-glutamic acid is produced and accumulated in a culture medium by culturing a mutant designated as FERM P-12379 which is derived from Escherichia coli K-12 strain and the mutant is deficient or low in .alpha.-ketoglutaric acid dehydrogenase activity, has low L-glutamic acid decomposing ability, and is capable of producing L-glutamic acid.

Abstract: The present invention provides a mutant and a process for producing L-glutamic acid by fermentation using a microorganism belonging to the genus Escherichia. In the present process, L-glutamic acid is produced and accumulated in a culture medium by (A) culturing an Escherichia mutant which is deficient or low in .alpha.-ketoglutaric acid dehydrogenase activity, has low L-glutamic acid decomposing ability, and is capable of producing L-glutamic acid.

Abstract: In a method for early diagnosis of human cancer, a human fecal sample of bacteria (Escherichia coli and/or Streptococcus faecalis), is incubated in vitro with a standard culture of a known number of cancer cells, for a period of time sufficient to enable the extent of interaction between the bacteria and the standard culture of cancer cells to be determined; the number of the interacted and/or non-interacted cancer cells present at the end of the period is determined and is utilized for the diagnosis based on the calculation of a tumor cell necrosis index (TCNI). The extent of interaction referred to may be calibrated against analogous interaction using a control preparation of bacteria, e.g. Escherichia coli A.T.C.C. 55373, 55374 and/or 55375, and/or Streptococcus faecalis A.T.C.C. 55376.

Abstract: The present invention relates to a method for identifying a TonB inhibitor in a test sample comprising:(a) growing a TonB.sup.+ microorganism in the presence of the test sample and a lethal agent, the activity of which is mediated by TonB;(b) identifying as positive a test sample with which the lethal activity of the agent is not observed;(c) growing on a low-iron medium a TonB.sup.+ microorganism in the presence of the test sample identified as positive in (b);(d) confirming as positive a test sample with which growth inhibition of the microorganism is observed on a low-iron medium.

Abstract: A rapid process for detecting pathogenic microorganisms in products for human consumption comprises contacting the microorganisms with a methylumbelliferone substrate. The substrate is hydrolyzed into methylumbelliferone by an enzyme given off by the microorganisms. Hydrolysis is accelerated by sodium lauryl sulfate, which renders the microorganisms more permeable to the substrate, the enzyme, or both. The methylumbelliferone is detected by its fluorescence, either in solution or on an agar medium supporting microcolonies formed from individual microorganisms.

Abstract: Microorganisms belonging to the genus Providencia or the genus Escherichia and having a resistance to isoleucine antagonist, produce L-threonine by fermentation in higher yield and in more amount of L-threonine accumulated.

Abstract: The Specification discloses:1. Recombinant microbial cloning vehicles comprising heterologous DNA coding for the expression of mammalian hormone (e.g., somatostatin) and other polypeptides, including plasmids suited for the transformation of bacterial hosts. The latter incorporate a regulon homologous to the host in its untransformed state, in reading phase with the structural gene for the heterologous DNA;2. Cloning vehicles coding for the microbial expression of a protein variously comprising (a) a polypeptide hapten and additional protein sufficient in size to confer immunogenicity on the product of expression, which may find use in raising antibodies to the hapten for assay use or in the manufacture of vaccines; and (b) a desired polypeptide product and additional protein from which the desired product may be cleaved; and3. Methods of preparing synthetic structural genes coding for the expression of mammalian polypeptides in microbial cloning systems.

Abstract: E. coli mutants which are capable of expressing cloned proteins in a highly stable manner are described.

Abstract: A novel hybrid B.t. toxin gene toxic to lepidopteran insects has been cloned. The DNA encoding the B.t. toxin can be used to transform various prokaryotic and eukaryotic microbes to express the B.t. toxin. These recombinant microbes can be used to control lepidopteran insects in various environments.

Abstract: The invention relates to a fragment of DNA containing genes encoding the synthesis of the O-antigen of Vibrio cholerae serotypes Inaba or Ogawa and being at least 16 kb in length. The invention further related to a cosmid comprising a cloned DNA fragment containing genes encoding the synthesis of O-antigen of Vibrio cholerae serotypes Inaba or Ogawa and to a strain of E.coli that includes the fragment.

Abstract: A novel B.t. toxin gene encoding a protein toxic to lepidopteran insects has been cloned from a novel lepidopteran-active B. thuringiensis microbe. The DNA encoding the B.t. toxin can be used to transform various prokaryotic and eukaryotic microbes to express the B.t. toxin. These recombinant microbes can be used to control lepidopteran insects in various environments.

Abstract: Novel plasmid vectors are described, for expression in Escherichia coli and/or Bacillus subtilis, in which the gene which codes for a heterologous protein is placed under the control of a promoter of the erythromycin gene which permit the organisms transformed with the abovementioned plasmids to express the heterologous protein in high yields.

Abstract: DNA sequences are provided coding for Bacillus thuringiensis var. israelensis (BTI) endotoxin, employing bacterial hosts which produce a protein having insecticidal activity for dipteran insects.The bacteriophage lambda strain SYN A4-1 was deposited at the A.T.C.C. on Feb. 22, 1984, and given Accession No. 40098.

Abstract: Antibodies are produced by hyperimmunizing a mammal, such as cow, with a vaccine derived from E. coli bacteria. The bacterial strains in the vaccine are selected on the basis of their virulence characteristics, especially adhesion factors (pili), associated with gastroenteric disease in humans. The antibodies can be recovered from the mammal's milk or serum, and used in human foods.

Abstract: The toxin gene encoding a protein toxic to beetles of the order Coleoptera, named M-7, has been cloned and expressed. M-7 is a novel Bacillus thuringiensis strain which has been deposited with a recognized culture repository. The microbe is now known as B. thuringiensis strain san diego.

Abstract: A biologically engineered plasmid coding for the production of .beta.-glucosidase. The plasmid can be incorporated into various microorganisms to enable the microorganism to digest cellobiose, which is produced from cellulose by an endo- or exocellular cellulase. One particular application is the incorporation of this plasmid into a microorganism which produces ethanol. Preferably, this ethanol producing microorganism is also ethanol tolerant.

Abstract: An L-glutamic acid producing microorganism, which is obtained by incorporation into a host strain of the genus Escherichia of a hybrid plasmid having inserted therein a DNA fragment with genetic information controlling L-glutamic acid production, said fragment being derived from a donor strain of Escherichia which is capable of producing L-glutamic acid useful for the production of high levels of L-glutamic acid.

Abstract: An L-valine-producing microorganism which is constructed by incorporation into a host strain of the genus Escherichia of a hybrid plasmid having inserted therein a DNA fragment with genetic information related to L-valine production which is derived from a donor strain of the genus Escherichia which is resistant to a valine analogue, is useful for the production of high levels of L-valine by fermentation.

Abstract: A microorganism of the genus Escherichia incorporated with a hybrid plasmid, which have been inserted with a DNA fragment possessing genetic information related to L-histidine production and obtained from a mutant of the genus Escherichia, resistant to a histidine-analogue, produces L-histidine in a high yield.

Abstract: The present invention relates to a process for the production of human erythropoietin.More precisely, the invention relates to a process for the mass production of human erythropoietin, comprising in vivo multiplication of human lymphoblastoid cells capable of producing human erythropoietin, and human erythropoietin production by the multiplied human lymphoblastoid cells.The human erythropoietin production according to the present invention is much higher, in terms of human erythropoietin production per cell, than that attained by conventional processes using in vitro tissue culture; thus, human erythropoietin can be used in a sufficient amount for the prevention and treatment of human diseases.

Abstract: A streptomycin dependent mutant of a microorganism of the genus Escherichia which contains a plasmid containing genetic information controlling streptomycin independence maintains its properties when cultured in a medium devoid of streptomycin. The plasmid may also contain genetic information controlling the production of a chemical compound by the microorganism. Fermentation cultures of such microorganisms in media devoid of streptomycin do not lose their industrially desirable ability to synthesize useful compounds.

Abstract: DNA comprising the naturally occurring nucleotide sequence coding for amino acids 44-90 of .beta.-lipotropin and including the entire coding region for .beta.-endorphin with the exception of the C-terminal glutamine was modified, transferred to an expression transfer vector, and expressed as a fusion protein. The fusion protein was further modified in vitro to yield mature .beta.-endorphin. .beta.-endorphin was purified from a bacterial lysate. The structure and biological activity of the resulting product was proven by immunological assay, and by two independent assays designed to demonstrate biological activity.

Abstract: Novel chemical compounds, cointegrate plasmids pUC1012 and pUC1013, which are obtained by covalent linkage of the E. coli plasmid pBR322 to the Streptomyces espinosus plasmid pUC6, and plasmids pUC1015 and pUC1022 which are obtained by restructuring plasmid pUC1012, and plasmids pUC1016 and pUC1023 which are obtained by restructuring plasmid pUC1013. These plasmids are useful as cloning vehicles in recombinant DNA work. For example, using DNA methodology, a desired gene, for example, the insulin gene, can be inserted into the plasmids and the resulting plasmids can then be transformed into a suitable host microbe which, upon culturing, produces the desired insulin.

Abstract: Novel chemical compounds, recombinant plasmids pUC1026 and pUC1027, which are obtained by covalent linkage of the E. coli plasmid pBR322 to the Streptomyces espinosus plasmid pUC6. These plasmids are produced by a novel process which can be used to stabilize unstable potential plasmid vectors. These plasmids are useful as cloning vehicles in recombinant DNA work. For example, using DNA methodology, a desired gene, for example, the insulin gene, can be inserted into the plasmids and the resulting plasmids can then be transformed into a suitable host microbe which, upon culturing, produces the desired insulin. The stabilization process disclosed herein can be used to make other stable plasmids.

Abstract: Disclosed are optically active acylated cephalosporin analogs which are useful as antibacterial agents and methods for preparing such compounds.

Abstract: Disclosed are optically active cephalosporin analogs which are produced by optically selective deacylation of an optically inactive acylated analog. The compounds are useful as intermediates in the preparation of optically active acylated antimicrobial agents.