Abstract: Described herein are methods and apparatus for assays of bacterial spores. In particular, methods and apparatus for lateral flow immunoassay for bacterial spore detection and quantification, live/dead assay for bacterial spores, lifetime-gated measurements of bacterial spores and imaging bacterial spores using an active pixel sensor, and unattended monitoring of bacterial spores in the air are described.

Abstract: Compositions and methods of killing, inactivating, or otherwise reducing the spores such as bacterial spores are disclosed. The methods typically include reducing or preventing spore reactivation comprising contacting spores with an effective amount of one or more green tea polyphenols (GTP), one or more modified green tea polyphenols (LTP), or a combination thereof. In a preferred embodiment, the LTP is (?)-epigallocatechin-3-gallate (EGCG) esterified at the 4? position with stearic acid, EGCG esterified at the 4? position with palmitic acid, or a combination thereof. The compositions and methods can be used in a variety of applications, for example, to increase the shelf-life of a food or a foodstuff, to reduce or delay the spoilage of a food or a foodstuff, or to decontaminate a device contaminated with spores.

Abstract: The present application relates to a method for inducing germination of a spore forming bacterium, specifically, a method for inducing germination of a spore forming bacterium, comprising: using one or more compounds having a phenolic backbone to induce germination of a spore forming bacterium selected from the group consisting of the genus Bacillus, the genus Geobacillus, and the genus Aeribacillus; to a germination inducer for a spore forming bacterium; and to a kit for use in germination of a spore forming bacterium.

Abstract: The process for the production of a chlamydospore rich slurry inoculum begins with a substrate colonized with a desired Basidiomycete fungus capable of producing chlamydospores during vegetative growth. The colonized substrate is treated to increase the chlamydospore production and content in said spawn and thereafter combined with water at rate of at least 1:6 spawn:water to obtain a slurry inoculum. The inoculum may then be agitated to populate a water fraction with chlamydospores or macerated to homogenously distribute the chlamydospores.

Abstract: A biosorbent for removing cationic and/or anionic metals from aqueous solutions, and a process for the production of the biosorbent. The biosorbent includes bacterial aggregates of Bacillus sp. VCHB-10, deposited as NRRL-B-30881, and treated with polyethyleneimine and glutaraldehyde. Among the metals in their cationic form, the following are considered: cations of Ag, Al, Au, Co, Cd, Cu, Cr, Fe, Hg, Mn, Ni, Pb, Pd, Pt, U, Th and Zn. Among the metals in their anionic form, the following are considered: anions of As, Cr and Mo. Removal or recovery of metals from wastewater using the biosorbent is also described.

Abstract: The present invention is a method for preparing a biocontrol formulation by the following steps. A Streptomyces bacterium is selected simultaneously with superior sporulation ability, chitinase activity and antagonistic effectiveness against plant fungal pathogens by culturing with fungal pathogens, wherein the selected Streptomyces bacterium is cultured on a chitin-limited plate to assess the ability of sporulation and chitinase activity, and the Streptomyces bacterium with superior sporulation ability is the strain with producing powder-like and maturity form spore chains. Spores from the selected bacterium is collected. The obtained spores are taken as a seed inoculum. Then the seed inoculum is cultured to directly yield a broth medium containing at least 109 viable spores/ml which the spores contained in the broth medium are well suspended. The biocontrol formulation is obtained. The present invention further provided a biocontrol formulation containing a high concentration of Streptomyces spp. spores.

Abstract: Current approaches for treating cancer are limited, in part, by the inability of drugs to affect the poorly vascularized regions of tumors. We have found that spores of anaerobic bacteria in combination with agents which interact with microtubules can cause the destruction of both the vascular and avascular compartments of tumors. Two classes of microtubule inhibitors were found to exert markedly different effects. Some agents that inhibited microtubule synthesis, such as vinorelbine, caused rapid, massive hemorrhagic necrosis when used in combination with spores. In contrast, agents that stabilized microtubules, such as the taxane, docetaxel, resulted in slow tumor regressions that killed most neoplastic cells. Remaining cells in the poorly perfused regions of tumors could be eradicated by sporulated bacteria.

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: A process is provided that is effective for allowing bacteria to be cultured in a selected substrate. Bacteria are sporulated and then germinated in the presence of a selected substrate and medium.

Abstract: Subpopulations of spore-like cells expressing specific cell surface and gene expression markers are provided. In one embodiment, the cells express at least one cell surface or gene expression marker selected from the group consisting of Oct4, nanog, Zfp296, cripto, Gdf3, UtF1, Ecat1, Esg1, Sox2, Pax6, nestin, SCA-1, CD29, CD34, CD90, B1 integrin, cKit, SP-C, CC10, SF1, DAX1, and SCG10. Also provided are methods for purifying a subpopulation of spore-like cells of interest from a population of spore-like cells, and methods for inducing differentiation of the isolated spore-like cells into cells of endodermal, mesodermal or ectodermal origin. The spore-like cells can be used to generate cells originating from all three germ layers and can be used to treat a patient who has a deficiency of functional cells in any of a wide variety of tissues, including the retina, intestine, bladder, kidney, liver, lung, nervous system, or endocrine system.

Abstract: A method of synthesizing optically-active (S)-3-hydroxybutyric acid and (S)-3-hydroxybutyrate ester using a mutated microorganism is provided. More particularly, a mutated microorganism for preparing (S)-3-hydroxybutyric acid transformed with a gene encoding ?-ketothiolase, a gene encoding (S)-3-hydroxybutyryl CoA dehydrogenase and a gene encoding acyl CoA hydrolase; a method of preparing (S)-3-hydroxybutyric acid using the mutated microorganism; a mutated microorganism for preparing (S)-3-hydroxybutyrate ester transformed with a gene encoding ?-ketothiolase, a gene encoding (S)-3-hydroxybutyryl CoA dehydrogenase, a gene encoding acyl CoA hydrolase and a gene encoding lipase; and a method of preparing (S)-3-hydroxybutyrate ester using the mutated microorganism are provided.

Abstract: A method for controlling Dipteran larvae or a method for inhibiting the development of larvicidal resistance, controlling resistant populations and reducing resistance levels in Diptera by introducing a larvicidally-effective amount of a combination of a strain of Bacillus thuringiensis subspecies israelensis and a strain of Bacillus sphaericus into an environment containing Dipteran larvae; and a composition of the combination are disclosed. Preferably both strains are non-genetically modified.

Abstract: The method disclosed herein, relates generally to introducing molecules such as biomolecules (e.g., nucleic acids) into a filamentous fungus. More specifically, the methods disclosed herein relate to introducing one or more nucleic acids into a filamentous fungus.

Abstract: The presently-disclosed subject matter is directed to biosensors comprising spore-forming bacterial cells and/or spores generated therefrom, a recognition unit within each spore-forming cell for binding an analyte of interest, and a reporter molecule within each spore-forming cell for detecting binding of the analyte of interest, wherein the reporter molecule generates a detectable signal upon binding of the analyte by the recognition element. The presently-disclosed subject matter further provides methods of using the biosensors and systems and kits including the biosensors.

Abstract: Filamentous fungi are grown in pellet form by culturing the filamentous fungi in liquid culture under one or more of the following conditions: 1) with addition of particulate substrates: 2) using spores which have been stored for a period of time prior to inoculation; and 3) using high spore inoculum concentrations.

Abstract: The invention provides a method for isolating and culturing a previously unculturable microorganism, which comprises: (i) collecting a sample from an environmental source; (ii) counting/estimating the number of microorganisms in the sample; (iii) diluting the sample in an appropriate medium; (iv) adding a gelating agent such as to entrap one or more microorganisms within a sphere of the gelating agent; (v) coating the spheres containing the entrapped microorganism(s) with a natural or synthetic polymer to form a polymeric membrane; (vi) incubating the coated spheres in the original environment for an appropriate time; (vii) cutting the spheres and scanning for microorganisms colonies; and (viii) isolating the microorganisms, and repeating steps (iii) to (vii) until a pure clone of said previously unculturable microorganism is obtained.

Abstract: A method for isolating microorganisms from a sample, the sample including sample matrix and microorganisms, the method including the steps of providing a receptacle, the receptacle configured to allow filtering of the sample and to reversibly contain the sample and a concentration agent; adding the sample to the receptacle, wherein a microorganism-bound composition will be formed in the receptacle, the microorganism-bound composition including concentration agent-bound microorganisms and sample matrix; and filtering the microorganism-bound composition through a filter to collect the concentration agent-bound microorganisms on the filter, wherein the filter has an average pore size that is greater than the average size of the microorganisms. Kits and systems are also disclosed herein.

Abstract: Disclosed are quick dissolve tablets, each including Lactobacillus delbrueckii subsp bulgaricus (LBD) and N-acetyl-glucosamine (NAG), as well as excipients, for oral mucosal administration, for improving the quality of life of Hepatitis C patients. Any formulation suitable for oral mucosal administration can be employed for administering the active ingredients in a sufficient dosage for therapeutic effect, one such formulation being: 50 mg of Lactobacillus delbrueckii subsp bulgaricus lysate strain YB-I 10 mg of N-acetyl D-glucosamine. Excipients can include one or more of, maltodextrin; xanthan gum; acesulfam K; lemon powder and a flavoring, e.g., juice; Mannitol TL-32-04, Microcrystalline Cellulose and Carrageenan, Fructose, PVP-XL TL-11-04, Gellan Gum, Citrus TL 1-04, Orange TL 19-04, Sucrolose TL-13-04, and Mg ST TL-13-04.

Abstract: Current approaches for treating cancer are limited, in part, by the inability of drugs to affect the poorly vascularized regions of tumors. We have found that spores of anaerobic bacteria in combination with agents which interact with microtubules can cause the destruction of both the vascular and avascular compartments of tumors. Two classes of microtubule inhibitors were found to exert markedly different effects. Some agents that inhibited microtubule synthesis, such as vinorelbine, caused rapid, massive hemorrhagic necrosis when used in combination with spores. In contrast, agents that stabilized microtubules, such as the taxane docetaxel, resulted in slow tumor regressions that killed most neoplastic cells. Remaining cells in the poorly perfused regions of tumors could be eradicated by sponzlated bacteria.

Abstract: Methods and devices for the interfacing of microchips to various types of modules are disclosed. The technology disclosed can be used as sample preparation and analysis systems for various applications, such as DNA sequencing and genotyping, proteomics, pathogen detection, diagnostics and biodefense.

Abstract: The invention provides methods and related products for extracting nucleic acids such as DNA from prokaryotic spores. The invention also encompasses methods for identifying the source of such spores via analysis of the isolated nucleic acids.

Abstract: Provided is a method for mapping traits in organisms, in particular in plants. The method comprises a) providing a population of SDR-0 organisms, in particular plants, that each arise from one member of a population of unreduced cells resulting from second division restitution, in particular a population of unreduced spores; b) producing SDR-1 progeny populations of each of these SDR-0 organisms; c) phenotyping the SDR-1 progeny populations to identify segregating traits within each SDR-1 progeny population; d) if segregating progeny are present in a SDR-1 progeny population, genotyping the corresponding SDR-0 organism and comparing the genotype thereof with the genotype of the other SDR-0 organisms to identify heterozygous chromosomal regions associated with the occurrence of the segregating trait identified in the SDR-1 progeny population.

Abstract: A method for the simultaneous concentration of multiple toxins from large volumes of water. The method includes the steps of providing a disposable separation centrifuge bowl, the centrifuge bowl including a positively charged material at it's inner core. A large water sample contaminated with toxins from a group consisting of protozoa, bacteria, bacterial spores, and toxins is delivered to the centrifuge bowl. A centrifugal force is applied to the separation bowl. The water sample is concentrated to remove large particles of the toxins in the bowl due to the centrifugal forces. The concentrated water sample is passes through the positively charged inner core to capture any remaining concentrated targets by electrostatic forces and the concentrated targets are eluted.

Abstract: Filamentous fungi are grown in pellet form by culturing the filamentous fungi in liquid culture under one or more of the following conditions: 1) with addition of particulate substrates: 2) using spores which have been stored for a period of time prior to inoculation; and 3) using high spore inoculum concentrations.

Abstract: The present invention provides a method for making pressure-induced ganoderma spores, for using the pressure-induced ganoderma spores to ameliorate and/or prevent influenza in a mammal; and to stimulating the host immune system in a mammal. The method for making the pressure-induced ganoderma spores includes (1) soaking ganoderma spores in a solution; (2) drying the soaked ganoderma spores using a freeze-drying or a vacuum-drying method; (3) breaking the sporoderms of the dried ganoderma spores with enzyme or a mechanical method; (4) pressurizing and depressurizing the sporoderm-broken ganoderma spores to obtain the pressure-induced sporoderm-broken ganoderma spores. The methods for ameliorating and/or preventing influenza, and for stimulating the host immune system include orally administering a composition comprising the pressure-induced ganoderma spores to a mammal susceptible to or suffering from influenza. The present invention also provides the pressure-induced ganoderma spores.

Abstract: An integrated industrial plant includes various systems, all of which use a cryogenic liquid obtained from a common source. One system includes a fermentation unit, in which cold air, chilled by heat exchange with the cryogenic liquid, absorbs excess heat generated by the fermentation. Another system is a lyophilization unit, in which a refrigeration step is performed through the use of air that has been chilled by heat exchange with the cryogenic liquid. Another system is a device for freezing discrete samples of biological products, the samples being frozen by partial immersion in the cryogenic liquid. The invention substantially reduces the use of electric power, and provides systems which operate economically and reliably.

Abstract: A method of detecting the presence and quantity of bacterial spores, which includes adding an electrophilic alcohol and an acid anhydride to a sample, admixing the sample with a solvent, and analyzing the sample. The sample may be analyzed by injecting the mixture into a gas chromatograph equipped with a mass spectra detector.

Abstract: The invention concerns an acellular immunogenic or vaccine composition for producing antibodies against Bacillus anthracis comprising a protective antigen (PA) and killed and optionally purified spores, obtained from mutating strains of Bacillus anthracis and their uses.

Abstract: The present invention provides a process for efficiently producing sporangia of Bacillus popilliae containing spores and parasporal bodies having controlling effects on Scarabaeidae insects, and a control agent and controlling method for Scarabaeidae insects obtained by said production process. In a process for producing sporangia of Bacillus popilliae containing spores and parasporal bodies by culturing Bacillus popilliae in a medium containing an adsorbent, the medium contains 0.2-4.0% by weight of glutamic acid.

Abstract: The present invention provides improved methods and compositions for producing oocysts. The oocysts produced according to the invention find use in the manufacture of vaccines. In preferred embodiments, the present invention provides methods and compositions for the production of Eimeria oocysts. Vaccines containing Eimeria oocysts, sporocysts and/or sporozoites produced according to the present invention may be used to immunize birds against coccidiosis either in ovo or post hatch.

Abstract: The present invention provides a process for producing sporangia of Bacillus popilliae containing spores and parasporal bodies in large numbers per unit volume of medium. In a process for producing sporangia of Bacillus popilliae containing spores and parasporal bodies by culturing Bacillus popilliae in a liquid medium containing an adsorbent, the liquid medium contains 0.1–0.7% by weight of proline.

Abstract: The invention relates to a method of vaccinating a domesticated bird against coccidiosis comprising administering in ovo an effective immnunizing dose of live Eimeria sporozoites or merozoites, or a mixture thereof. In a preferred embodiment, the domesticated bird that is vaccinated is a chicken or turkey.

Abstract: The invention concerns an acellular immunogenic or vaccine composition for producing antibodies against Bacillus anthracis comprising a protective antigen (PA) and killed and optionally purified spores, obtained from mutating strains of Bacillus anthracis and their uses.

Abstract: A process to excystate protozoal sporocysts involves treatment of an infected tissue sample with a sodium hypochlorite solution to stimulate excystation of the sporocysts from the tissue. Thereafter, removal of the sodium hypochlorite solution and treatment of the sample with a cell culture media as the excystation fluid eliminates incubation and subsequent washing steps using expensive reagents. The excystation fluid contains substantially no chelating agents, proteins, enzymes or bile acids.

Abstract: Methods and devices are provided for the detection of bacterial agents such agents as Bacillus anthracis and Clostridium botulinum with high sensitivity and selectivity. More specifically, methods and devices are based on a phosphorescence-emission detection system using chelate-stabilized lanthanides (e.g, Eu(III), Tb(III), and Sm(III)) to detect various spore-specific small organic molecules (e.g., dipicolinic acid, diaminopimelic acid, n-acetlymuramic acid, and the like). By careful selection of the chelating agent or ligand coordinated to the lanthanide, both high specificity and selectivity can be obtained. Examples of suitable and preferred sensor systems include N-(2-hydroxyethyl)ethylenediaminetriacetic acid (HEDTA) and N-(2-hydroxyethyl)iminodiacetic acid (HEIDA) combined with europium (III) and/or terbium (III). The chelate-stabilized lanthanides react with the spore-specific “target” molecules to form a characteristically phosphorescent product which can then be detected.

Abstract: A process for structuring a surface layer of an object includes applying bio-components to the surface of the object that carry away surface material. The bio-components are contained in at least one of a nutrient and osmotic protective medium. The at least one of a nutrient and osmotic protective medium having the bio-components contained therein is removed after the surface material is carried away from the object surface.

Abstract: This invention relates to a process for detecting the presence of viable bacterial spores in a sample and to a spore detection system, the process including placing a sample in a germination medium for a period of time sufficient for commitment of any present viable bacterial spores to occur, mixing the sample with a solution of a lanthanide capable of forming a fluorescent complex with dipicolinic acid, and, measuring the sample for the presence of dipicolinic acid, and the system including a germination chamber having inlets from a sample chamber, a germinant chamber and a bleach chamber, the germination chamber further including an outlet through a filtering means, the outlet connected to a detection chamber, the detection chamber having an inlet from a fluorescence promoting metal chamber and the detection chamber including a spectral excitation source and a means of measuring emission spectra from a sample, the detection chamber further connected to a waste chamber.

Abstract: The invention relates to a method of vaccinating a domesticated bird against coccidiosis comprising administering in ovo an effective immunizing dose of live Eimeria sporocysts or oocysts, or a mixture thereof.

Abstract: A strain of Trichoderma harzianum is obtained that is useful as a nematode inhibitor, fungicide and plant growth promoter. The strain has ATCC accession number PTA-3701. The strain is isolated by treating Trichoderma harzianum isolated from experimental fields of Central Institute of Medicinal and Aromatic Plants (CIMAP) Field Station with a mutagen such as ethyl methyl sulphonate, and isolating a whitish and fast growing strain of Trichoderma harzianum.

Abstract: A method is disclosed for determining the viability of sporocyst-forming protozoa. The method involves treating a sample of protozoa with at least one vital dye and determining the viability of the protozoa in the sample by differential staining. The viability of protozoa in the sample can then be correlated with the viability of protozoa in the population from which the sample was obtained.

Abstract: The invention provides def1 polypeptides and polynucleotides encoding def1 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing def1 polypeptides to screen for antibacterial compounds.

Abstract: Embryogenesis from plant microspores is routinely induced with a 16-24 h temperature treatment of 32.5° C. Continuous culture at 25° C. results in pollen development. However, microspore treatment with anti-cytoskeletal agents, or protein synthesis inhibitors, at the non-inductive temperature of 25° C., can induce embryogenesis, thus demonstrating that heat shock is not required for embryogenic induction. Furthermore, when anti-microtubule agents (e.g. colchicine) are used, embryo induction and chromosome doubling occur simultaneously, thus generating doubled haploids, whereas heat induction generates haploids. Thus, the use of microtubule inhibitors will provide a simple one-step process to simultaneously induce embryogenesis and chromosome doubling for the production of fertile plants, thus providing minimal manipulation which will be very advantageous for genetic studies and plant breeding programs. As noted, heat shock induces haploids.

Abstract: A fungal inoculum comprising a pelleted nutrient substrate coated with fungal propagules suspended in a hydrophilic material is disclosed.

Abstract: Embryogenesis from plant microspores is routinely induced with a 16-24 h temperature treatment of 32.5.degree. C. Continuous culture at 25.degree. C. results in pollen development. However, microspore treatment with anti-cytoskeletal agents, or protein synthesis inhibitors, at the non-inductive temperature of 25.degree. C., can induce embryogenesis, thus demonstrating that heat shock is not required for embryogenic induction. Furthermore, when anti-microtubule agents (e.g. colchicine) are used, embryo induction and chromosome doubling occur simultaneously, thus generating doubled haploids, whereas heat induction generates haploids. Thus, the use of microtubule inhibitors will provide a simple one-step process to simultaneously induce embryogenesis and chromosome doubling for the production of fertile plants, thus providing minimal manipulation which will be very advantageous for genetic studies and plant breeding programs. As noted, heat shock induces haploids.

Abstract: The invented methods are effective for the detection and quantification of bacterial spores in a sample medium. A lanthanide such as europium or terbium is combined with a medium to be tested for endospore content. The lanthanide will react with calcium dipicolinate present in any bacterial spores in the sample medium to produce a lanthanide chelate, specifically, terbium or europium dipicolinate. The lanthanide chelate has distinctive absorbance and emission spectrums that can be detected using photoluminescence testing, for example. The occurrence of emission from the sample medium upon excitation at wavelengths distinctive of the lanthanide chelate thus reveals the presence of spores in the sample medium. The concentration of spores can be determined by preparing a calibration curve that relates absorbance or emission intensities to spore concentrations for test samples with known spore concentrations.

Abstract: The present invention relates to a novel sporulation gene isolated from B. thuringiensis, and a DNA segment comprising the nucleic acid sequences encoding the gene and DNA sequences encoding crystal toxin proteins wherein said DNA segment is stably integrated into a B. thuringiensis host by homologous recombination. The invention further relates to a DNA segment wherein the sporulation gene is mutated thereby rendering an oligosporogenic or asporogenic transformed B. thuringiensis as a result of stable integration of the DNA segment into the Bacillus thuringiensis chromosome by homologous recombination.

Abstract: A bacteria impermeable container or ampule (10) contains a liquid growth medium and a substrate-indicator complex. The complex includes a substrate component, e.g., starch, and an indicator molecule, e.g., a dye, a fluorescent molecule, or the like, which are tightly bound and complexed, but which are cleavable by a preselected enzyme. A sterilant passes over a carrier (20) for microorganisms which, upon germination, are capable of rapidly generating large quantities of the preselected enzyme. Following the sterilization process, the carrier is immersed in the liquid growth medium. Any viable surviving microorganisms grow, generating the preselected enzyme. The enzymes cleave the bound indicator molecule from the substrate, resulting in a measurable property change in a couple of hours. Typical property changes include fluorescence, a color change, a change in pH which triggers a pH indicator color change, and the like.

Abstract: The invention provides protein-based sensors which report changes at a cell surface membrane. These sensors comprise an endogenous cell surface protein having a post-translationally generated luminescer at a predetermined residue. In operation, the protein adopts one of a plurality of different interconvertable signal-dependent conformations, whereunder the luminescer provides corresponding different luminescence. The cells comprise transgenes encoding the protein sensor which is translated from such transgenes by the host cell's machinery with natural amino acids and expressed on the cell surface. After surface expression, the cell is contacted with a luminescer generating reagent which generates a luminescer at a predetermined residue of the protein. A wide variety of luminescers and chemistries for generating the selected luminescer at the target residue of the sensor protein may be used.

Abstract: The present invention is directed to a family of phospholipid binding and transporting proteins, a method for preparing same from fungi, and their use in cosmeticology, the agri-foodstuffs industry and pharmacology Phospholipid proteins are capable of binding, transporting and/or rearranging lipids between membranes, optionally in combination with active principles. Furthermore, the phospholipid proteins are hydrophobic and acidic, have a molecular weight of under 50 kDa, and may be prepared from a non-toxic filamentous fungus capable of developing on a lipid-enriched medium, particularly from raw extracts of fungi.

Abstract: A method of determining the effectiveness of a cancer treatment by sealing tumor cells in segments of semipermeable membrane hollow fibers, implanting the sealed fiber segments in a mammal, treating the mammal with a cancer treatment, and evaluating the effect of the cancer treatment on the cells in the hollow fiber segments.