Abstract: Selective enrichment media and methods for selectively growing and detecting Salmonella spp. and/or Shiga toxin-producing E. coli. The media may comprise a carbon and nitrogen source, an inorganic salt, a fermentable sugar, one or more selective agents, and an efflux pump inhibitor. Various selective agents include sulfa drugs, surfactants, aminocoumarins, cycloheximide, supravital stains, ascorbic acid, bromobenzoic acid, myricetin, nitrofurantoin, rifamycins, polyketides, and oxazolidinones. Various efflux pump inhibitors include arylpiperazines, such as 1-(1-naphthylmethyl)piperazine, and quinoline derivatives, such as 4-chloroquinoline. Methods of selectively growing and detecting Salmonella and/or Shiga toxin-producing E. coli are provided.

Abstract: The present disclosure describes pathogenic bacteria that have been modified to be deficient in NiFe hydrogenase activity; compositions comprising such modified bacteria, and the use of such bacteria to protect animals from pathogenic enteric bacterial infections.

Abstract: Selective growth media N4 agars that include a combination of high levels of yeast extract, possibly higher levels of protein, elevated levels of sugar and reduced levels of sodium chloride. The combination of ingredients provides the ability to detect Salmonella spp. and Shigella spp. with almost identical sensitivity while alleviating false-negative and false-positive problems commonly encountered with the presence of Proteus spp. and Citrobacter spp.

Abstract: The present invention is directed to mutant Salmonella sp. having a genetically modified msbB gene in which the mutant Salmonella is capable of targeting solid tumors. The invention is also directed to Salmonella sp. containing a genetically modified msbB gene as well as an genetic modification in a biosynthetic pathway gene such as the purl gene. The present invention further relates to the therapeutic use of the mutant Salmonella for growth inhibition and/or reduction in volume of solid tumors.

Abstract: Monolithic bioelectronic devices for the detection of ammonia includes a microorganism that metabolizes ammonia and which harbors a lux gene fused with a heterologous promoter gene stably incorporated into the chromosome of the microorganism and an Optical Application Specific Integrated Circuit (OASIC). The microorganism is generally a bacterium.

Abstract: Attenuated immunogenic bacteria having an RpoS+ phenotype, in particular, Salmonella enterica serotype Typhi having an RpoS+ phenotype and methods therefor are disclosed. The Salmonella have in addition to an RpoS+ phenotype, an inactivating mutation in one or more genes which render the microbe attenuated, and a recombinant gene capable of expressing a desired protein. The Salmonella are attenuated and have high immunogenicity so that they can be used in vaccines and as delivery vehicles for genes and gene products. Also disclosed are methods for preparing the vaccine delivery vehicles.

Abstract: The present invention is directed to mutant Salmonella sp. having a genetically modified msbB gene in which the mutant Salmonella is capable of targeting solid tumors. The present invention further relates to the therapeutic use of the mutant Salmonella for growth inhibition and/or reduction in volume of solid tumors.

Abstract: The present invention is directed to mutant Salmonella sp. having a genetically modified msbB gene in which the mutant Salmonella is capable of targeting solid tumors. The invention is also directed to Salmonella sp. containing a genetically modified msbB gene as well as an genetic modification in a biosynthetic pathway gene such as the purI gene. The present invention further relates to the therapeutic use of the mutant Salmonella for growth inhibition and/or reduction in volume of solid tumors.

Abstract: A bacterial autoinducer and method for isolating and purifying a bacterial autoinducer form a sample comprising the steps of collecting a sample containing the autoinducer, fractionating the sample to isolate fractions corresponding to molecular weights of approximately 300-1500 Dalton, and eluting the isolate on an anion-exchange chromatographic column and selecting the faction containing the autoinducer.

Abstract: A method is provided for preserving live bacteria by subjecting an aqueous system containing the growing bacteria to drying without special equipment, in the presence of trehalose with or without the addition of divalent cations as stabilizing agents. Further, a dried composition for preservation of aerobic bacteria in a viable state is provided. The dried composition consists essentially of dried viable aerobic bacteria and an appropriate growth medium. The bacteria and growth medium are initially placed in an aqueous solution of 10 mM to 200 mM trehalose and a divalent cation, and dried at room temperature.

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: The present invention is directed to mutant Salmonella sp. having a genetically modified msbB gene in which the mutant Salmonella is capable of targeting solid tumors. The invention is also directed to Salmonella sp. containing a genetically modified msbB gene as well as an genetic modification in a biosynthetic pathway gene such as the purI gene. The present invention further relates to the therapeutic use of the mutant Salmonella for growth inhibition and/or reduction in volume of solid tumors.

Abstract: A method for vaccinating poultry to prevent salmonellosis and other microbial-related health problems in humans is described. The method involves isolation of a poultry heterophil-adapted strain of a microorganism that may be used in a vaccine. A vaccine comprising a preparation of the poultry heterophil-adapted strain is administered to poultry to reduce the transmission of microorganisms causing salmonellosis and other illnesses.

Abstract: A bacterial autoinducer and method for isolating and purifying a bacterial autoinducer from a sample comprising the steps of collecting a sample containing the autoinducer, fractionating the sample to isolate fractions corresponding to molecular weights of approximately 300-1500 Dalton, and eluting the isolate on an anion-exchange chromatographic column and selecting the faction containing the autoinducer.

Abstract: Attenuated Salmonella mutants which constitutively express the Vi antigen are disclosed, as well as vaccines against typhoid fever containing the same, live vector vaccines containing the same, and DNA-mediated vaccines containing the same.

Abstract: A method for vaccinating poultry to prevent salmonellosis and other microbial-related health problems in humans is described. The method involves isolation of a poultry heterophil-adapted strain of a microorganism that may be used in a vaccine. A vaccine comprising a preparation of the poultry heterophil-adapted strain is administered to poultry to reduce the transmission of microorganisms causing salmonellosis and other illnesses.

Abstract: This invention provides immunogenic compositions for the immunization of a vertebrate or invertebrate comprising an avirulent derivative of S. typhi. The derivatives having a mutation of the cya and/or crp and/or cdt genes. The invention also provides immunogenic compositions for the immunization of a vertebrate and invertebrate comprising an avirulent derivative of the above type which is capable of expressing a recombinant gene derived from a pathogen of said vertebrate or invertebrate individual to produce an antigen capable of inducing an immune response against said pathogen. Other embodiments of the invention include methods of preparing immunogenic compositions from these strains, and strains useful in the preparation of the immunogenic compositions, as well as methods of stimulating the immune system to respond to an immunogenic antigen of S. typhi by administration of the immunogenic composition.

Abstract: This invention provides immunogenic compositions for the immunization of a vertebrate or invertebrate comprising an avirulent derivative of S. typhi. The derivatives having a mutation of the cya and/or crp and/or cdt genes. The invention also provides immunogenic compositions for the immunization of a vertebrate and invertebrate comprising an avirulent derivative of the above type which is capable of expressing a recombinant gene derived from a pathogen of said vertebrate or invertebrate individual to produce an antigen capable of inducing an immune response against said pathogen. Other embodiments of the invention include methods of preparing immunogenic compositions from these strains, and strains useful in the preparation of the immunogenic compositions, as well as methods of stimulating the immune system to respond to an immunogenic antigen of S. typhi by administration of the immunogenic composition.

Abstract: A microbiological assay for chemicals, which uses a cell and a reducing dye to quantitatively measure inhibition of electron transport in the cell membrane as a function of chemicals in the substance being tested, is disclosed. This assay and method is reliable, simple, fast, and inexpensive, requires a minimum amount of durable equipment, and avoids the need for the use of live animals as the indicator organisms. The assay is particularly useful for testing for toxicity in food products, environmental, medical and industrial processes, sewage treatment, effluent, agricultural wastes, and chemical dumps.

Abstract: The invention comprises a culture medium and method for distinguishing bacteria of Salmonella species from other gram-negative bacteria, especially those belonging to the family Enterobacteriaceae. It is based on the ability of salmonellae to utilize melibiose, mannitol, and sorbitol and convert them into acids, together with a chromogenic substrate used for identifying .beta.-galactosidase. Other bacteria of the family Enterobacteriaceae, most of which are .beta.-galactosidase-positive, appear as brown, blue, or green colonies, depending on the chromogenic substrate used. Apart from Salmonella species, other .beta.-galactosidase-negative bacteria, such as Proteus species, appear as colorless colonies. Salmonellae can be identified directly on the culture medium after incubation, by their characteristic bright red color.

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 test method and medium for quantitatively identifying and distinguishing biological materials in a test sample. A first biological material has enzyme specificity for a first chromogenic substrate, a second biological material has enzyme specificity for a second chromogenic substrate, and a third biological material has specificity for one of the substrates. The chromogenic substrates form respective first and second colored water insoluble compounds upon reaction with specific enzymes. The first and second biological materials are capable of fermenting a sugar, and the third material does not ferment sugar. The test medium is adjusted to a pH conducive for color change of a pH indicator upon acidification due to fermentation, resulting in the formation of a zone of a third color around the water insoluble compounds of the sugar-fermenting materials.

Abstract: A novel method for conjugating haptens, ligands or luminescent labels to polynucleotides has been developed. This method involves the formation of a mixed anhydride, followed by reaction of the mixed anhydride intermediate with a nucleophilic group-containing polynucleotide in a DMF/water solvent system. The resulting compound finds usefulness especially in the gene probe area.

Abstract: Disclosed herein are methods for attenuating virulent gram negative bacteria to produce avirulent bacteria. The methods comprise passaging the wild-type bacteria through phagocytic cells, such as macrophages or polymorphonuclear leukocytes, or through lysosomes derived from such cells, a sufficient number of times until the bacteria become avirulent to the animal host. The bacteria are preferably from the family Enterobacteracea and most preferably from the genus Salmonellae. The invention further comprises the avirulent bacteria produced by the methods, pure cultures of such bacteria, and methods of using the bacteria, preferably in a vaccine for administration to an animal host to induce an immune response to the wild-type gram negative bacteria in the host.

Abstract: A method for testing causative bacterial species of food poisoning which is characterized by using two oligonucleotide primers that hybridize to opposite strands of bacterial DNA specifically, and flank a unique region in the target DNA and amplifying the specific fragment of the bacterial DNA, comprising the steps of:(a) hybridizing the primer to specific gene sequence of bacteria in a sample, extending the hybridized primer with deoxynucleotide triphosphates (dATP, dCTP, dGTP, and dTTP), and resultantly making the double strand nucleotide;(b) where the primer extension products are cleaved into each single strand of nucleotide by certain external force such as heat, pH and so on, one single strand functioning as a template for nucleotide extension with a primer of the other strand;(c) repeating a series of cycles involving cleavage of primer extension products, primer hybridizing, extension of the hybridized primers to amplify the specific fragment of DNA, and detecting the amplified DNA fragment; and(d) as

Abstract: Disclosed herein are methods for attenuating virulent gram negative bacteria to produce avirulent bacteria. The methods comprise passaging the wild-type bacteria through phagocytic cells, such as macrophages or polymorphonuclear leukocytes, or through lysosomes derived from such cells, a sufficient number of times until the bacteria become avirulent to the animal host. The bacteria are preferably from the family Enterobacteracea and most preferably from the genus Salmonellae. The invention further comprises the avirulent bacteria produced by the methods, pure cultures of such bacteria, and methods of using the bacteria, preferably in a vaccine for administration to an animal host to induce an immune response to the wild-type gram negative bacteria in the host.

Abstract: The invention described herein consists of a process for preparing an antigenic reagent useful for the indirect determination of Salmonella typhi, the organism that is the causal agent of typhoid fever (TF). The invention consists on the following steps: to grow Salmonella typhi in a culture medium, characterized by containing a free-iron chelator, which generates a specific S. typhi outer membrane protein (OMP) pattern, OMPs that are used as a selective antigen for the detection of specific serum antibodies, by an immunoassay technique (ELISA).

Abstract: This invention provides immunogenic compositions for the immunization of a vertebrate or invertebrate comprising an avirulent derivative of S. typhi. The derivatives having a mutation of the cya and/or crp and/or cdt genes. The invention also provides immunogenic compositions for the immunization of a vertebrate and invertebrate comprising an avirulent derivative of the above type which is capable of expressing a recombinant gene derived from a pathogen of said vertebrate or invertebrate individual to produce an antigen capable of inducing an immune response against said pathogen. Other embodiments of the invention include methods of preparing immunogenic compositions from these strains, and strains useful in the preparation of the immunogenic compositions, as well as methods of stimulating the immune system to respond to an immunogenic antigen of S. typhi by administration of the immunogenic composition.

Abstract: Cytokine receptors for tumor necrosis factor e which are found on microorganisms may, if bound with exogenous TNF.alpha., enhance the response of natural killer cells activated by the microorganisms, or increase TNF.alpha. production by peripheral blood lymphocytes treated with the microorganisms. Microorganisms with receptor-bound exogenous TNF.alpha. have enhanced cellular invasion ability which may change the immune response thereto. Clinical and pharmaceutical applications of these discoveries are provided.

Abstract: The invention relates to a method of detecting gram-negative bacterial endotoxin using antibody capture combined with amoebocyte lysate chromogenic detection. The method is highly sensitive and rapid and may be used for detection of specific endotoxin. In a particular application, picogram levels of Haemophilus influenzae type b endotoxin are detected in plasma taken from previously infected mammals. In another particular application, the method is applied to the detection and diagnosis of disease, through the detection of endotoxin from disease-causing organisms. A specific example is the diagnosis of chancroid through the detection of endotoxin from H. ducreyi.

Abstract: This invention provides immunogenic compositions for the immunization of a vertebrate or invertebrate comprising an avirulent derivative of S. typhi. The derivatives having a mutation of the cya and/or crp and/or cdt genes. The invention also provides immunogenic compositions for the immunization of a vertebrate and invertebrate comprising an avirulent derivative of the above type which is capable of expressing a recombinant gene derived from a pathogen of said vertebrate or invertebrate individual to produce an antigen capable of inducing an immune response against said pathogen. Other embodiments of the invention include methods of preparing immunogenic compositions from these strains, and strains useful in the preparation of the immunogenic compositions, as well as methods of stimulating the immune system to respond to an immunogenic antigen of S. typhi by administration of the immunogenic composition.

Abstract: Cytokine receptors for tumor necrosis factor .alpha. which are found on microorganisms may, if bound with exogenous TNF.alpha., enhance the response of natural killer cells activated by the microorganisms, or increase TNF.alpha. production by peripheral blood lymphocytes treated with the microorganisms. Microorganisms with receptor-bound exogenous TNF.alpha. have enhanced cellular invasion ability which may change the immune response thereto. Clinical and pharmaceutical applications of these discoveries including vaccines with increased efficacy are provided.

Abstract: A (1R,2S)-2-hydroxycycloalkanecarboxylic acid ester is efficiently and selectively produced by microbial asymmetric reduction of a 2-oxocycloalkanecarboxylic acid ester with a bacterial strain or its processed material.

Abstract: Live vaccines are provided and methods for preparing the live vaccines for protection of a host from a pathogenic microorganism. The vaccines are prepared by introducing at least one modification in a gene involved in at least one, normally at least two, biosynthetic pathways involving the production of products which are unlikely to be found in the disease susceptible host. The modification results in a gene change which cannot be repaired by a single step, e.g. polynucleotide deletions and inversions. Where the aro gene suffers such a change, the resultant auxotrophic mutants require aromatic amino acids, p-aminobenzoic acid and 2,3-dihydroxybenzoic acid or a highly concentrated source of absorabable iron. The auxotrophic mutations have substantially reduced or nonexistent virulence while retaining the desired immunogenicity to initiate the immunogenic response. Various techniques can be employed for providing the desired change.

Abstract: The invention relates to a method of detecting gram-negative bacterial endotoxin using antibody capture combined with amoebocyte lysate chromogenic detection. The method is highly sensitive and rapid and may be used for detection of specific endotoxin. In a particular application, picogram levels of Haemophilus influenzae are detected in plasma taken from previously infected mammals.

Abstract: The present invention relates to an isolating medium for the identification of the Salmonella bacterium, wherein a polyol metabolizable by Salmonella and a pH indicator reacting to acidification are added to a culture support containing peptones.

Abstract: Non-virulent aro-Salmonella typhimurium bacterium is disclosed into which has been cloned a heterologous nucleotide sequence encoding for the expression of Streptococcal M protein antigen from S. pyogenes seratype 5, which is effective to elicit opsonic antibodies against Streptococcal infections. The bacterium is useful for vaccination against Streptococcus pyogenes bacteria.

Abstract: Improved nucleic acid probes capable of specifically hybridizing to rRNA of Salmonella and not to rRNA of non-Salmonella are described along with methods utilizing such probes for the detection of Salmonella in food and other samples.

Abstract: The present invention is directed to attenuated strains of enteroinvasive bacteria that express a peptide or protein related to an epitope of the malaria parasites of the genus Plasmodium. The bacterial strains of the invention which can multiply in a host without causing significant disease or disorder, and which express a Plasmodium-related peptide that induces a protective immune response against malaria, can be used in live vaccine formulations for malaria. In specific embodiments, a Plasmodium-related peptide can be expressed as a fusion protein, for example, with a bacterial enterotoxin.The invention also relates to methods for expression of malaria antigens or fragments thereof within attenuated enteroinvasive bacteria.In particular embodiments, the invention is directed to the expression by attenuated Salmonella spp. of epitopes of Plasmodium circumsporozoite proteins.

Abstract: Nucleic acid probes capable of specifically hybridizing to rRNA of E. coli and Shigella species and not to rRNA of non-E. coli/Shigella are described along with methods utilizing such probes for the specific detection of E. coli and/or Shigella in food and other samples.

Abstract: Methods and compositions are provided for the cloning and expression of plasmids bearing genes coding for the non-toxic subunit of the heat-laible enterotoxin (LT-B) of E. coli. These plasmids may be cloned into stable avirulent strains of Salmonella typhi and used to make oral bivalent vaccines. Such vaccines may be used to prevent typhoid fever and cholera-like enterotoxin-induced diarrheal disease.

Abstract: This invention relates to a nucleic acid probe and method for the rapid detection of typhoid fever bacteria by use of a nucleic acid hybridization probe, equivalent to the DNA region encoding the Vi antigen of enteric bacteria such as Salmonella typhi, S. paratyphi C, or Citrobacter freundii, in a nucleic acid hybridization reaction with a clinical specimen containing typhoid fever bacteria.

Abstract: A novel probe for detecting Salmonella comprises:a labeled substance, anda DNA or RNA fragment which hybridizes with a base sequence having the following formula (I), or (II) being complementary to the formula (I).5'-GCTCAGACGTATGGCGGTA-3' (I)3'-CGAGTCTGCATACCGCCAT-5' (II)This invention also provides a method for detecting Salmonella by using the probe.The probe reduces the time required for detecting Salmonella.

Abstract: An agent for the diagnosis and treatment of malignant tumors and for the treatment of lowered cellular and humoral immune defense is described which contains an immune modulator, a lipopolysaccharide or an immune modulator tagged with a radioactive tracer, a dyestuff or a cytostatic, or a similar polysaccharide, possibly in and/or on liposomes or lipidized. There is also described a product which contains the agent together with an adjuvant consisting of an aldehyde and an alcohol. There is also described a method for the diagnosis of malignant tumors comprising the step of administering to a mammal a diagnostic effective amount of the agent, optionally together with an adjuvant consisting of an aldehyde and an alcohol.

Abstract: A method for rapidly and reliably determining the potential carcinogenic activity of hydrocarbon mixtures which is especially useful for those of petroleum origin. A sample of the mixture is nitrated under conditions effective to convert the polynuclear aromatic hydrocarbons to their nitrated derivatives, and without separation of the nitrated components, the product is incubated with an inoculum of Salmonella typhimurium tester strain T98. The excessive production of revertant colonies is a measure of the mutagenic activity of the oil, and this measure is shown to correlate with dermal carcinogenic activity.

Abstract: Methods and associated devices for detecting the presence of a particular motile organism within a sample are disclosed. The sample may be derived from dry milk, raw meat, poultry or a clinical specimen. A preferred method includes inoculating a selective enrichment medium containing a chemotactic attractant with the sample and contacting the selective enrichment medium with a nonselective motility medium containing a chemotatic attractant in a concentration less than the attractant concentration in the selective enrichment medium. Upon incubation, the motile organism metabolizes the chemotactic attractant, allowing the organism to move into the motility medium where it interacts with antibodies specific for the organism, thereby causing the formation of a persistent immobilization band. The methods are particularly useful in detecting Salmonella.

Abstract: Compost, e.g. hardwood bark, is rendered suppressive to plant pathogens, such as Rhizoctonia solani, Pythium ultimum and Fusarium, and/or diseases caused thereby by adding to the compost, desirably after peak heating has been achieved but before substantial recolonization of the compost by mesophilic microorganisms has occurred, one or more microorganisms antagonistic to the plant pathogen. Container media also is rendered suppressive to plant pathogens and/or diseases caused thereby by amending the media with the just-described prepared suppressive compost or, alternatively, by amending separately with the compost and with Trichoderma fungus and antagonistic bacterium separately or mixed together. Desirably, the inoculated antagonistic microorganisms comprise Trichoderma hamatum species A.T.C.C. No. 20765 or 20764, together with Xanthomonas maltophilia bacterium species A.T.C.C. No. 53199 or a Flavobacterium balustinum isolate 299, A.T.C.C. No. 53198 species, A.T.C.C. No. 53198.

Abstract: Live vaccines are provided and methods for preparing the live vaccines for protection of a host from a pathogenic microorganism. The vaccines are prepared by introducing at least one modification in a gene involved in at least one, normally at least two, biosynthetic pathways, involving the production of products which are unlikely to be found in the disease susceptible host. The modification results in a gene change which cannot be repaired by a single step e.g. polynucleotide deletions and inversions. Where the aro gene suffers such a change, the resultant auxotrophic mutants require aromatic amino acids, p-aminobenzoic acid and 2,3-dihydroxybenzoic acid or a highly concentrated source of absorbable iron. The auxotrophic mutations have substantially reduced or nonexistent virulence, while retaining the desired immunogenicity to initiate the immunogenic response. Various techniques can be employed for providing the desired change.Salmonella typhimurium strain SL1479 was deposited at the ATCC on Sept.

Abstract: A method of treating a human patient to effect the remission of symptoms associated with psoriasis, which comprises parenterally administering, in multiple injections, to the patient typhoid vaccine in a therapeutically effective amount which is sufficient to provide immunostimulating activity.

Abstract: L-phenylalanine is produced by using a microorganism belonging to the species Citrobacter freundii, Erwinia herbicola, Enterobacter cloacae, Klebsiella oxytoca, Salmonella typhimurium, Bacillus cereus, Flavobacterium suaveolens, Serratia marcescens, Pseudomonas putida, Enterobacter cloacae, Proteus mirabilis, Paracoccus denitrificans, Arthrobacter globiformis, Bacillus sphaericus, Corynebacterium hydrocarboclastus, Kluyvera micum or Microbacterium ammoniaphilum and having the ability to convert phenylpyruvic acid into L-phenylalanine in the presence of an amino group donor; or fumaric acid and ammonium ion or urea.