Abstract: This invention relates to a method for producing cells which are competent for transformation and which may be stably stored for extended periods of time at various temperatures. The method involves growing cells in a growth conducive medium, rendering said cells competent, and lyophilizing said competent cells. The invention further relates to competent cells produced by such a method, to methods of transforming said cells with a DNA molecule, and to a method of producing a desired protein or polypeptide from said transformed cells.

Abstract: The invention provides a scintillation proximity assay for detecting peptidoglycan synthesis. The assay is especially suitable for high throughput screening of compounds affecting peptidoglycan synthesis.

Abstract: A transport/preservation system has been found which provides safe, effective transport for patient specimens and bodily fluids, and functions as a preservative for biological reagents, therapeutics, and personal care products. The transport/preservative formulations can also be used as safe, effective sanitizers on surfaces, equipment, and appliances. The transport preservation formulation includes a biguanide and one or more other antimicrobial agents, which is cidal to microorganisms when present in a sample or on a surface to be sanitized.

Abstract: A method of rapid biochemical cycling of aquariums using naturally preserved granular marine substrate material, such as sand or aragonite, to rapidly denitrify the aquatic environment and to establish biochemical conditions that are favorable to the survival and viability of fish, crustaceans, invertebrates, and other marine aquatic life. The method includes the steps of harvesting and packaging marine sand such that marine microorganisms, in the form of biofilm attached to the sand, are preserved with an optimal amount of water and air in retail packaging specifically dimensioned and configured for maintaining ammonia oxidizing bacteria in a state wherein the bacteria are capable of metabolic and physiologic activity after prolonged periods at room temperature. Harvesting and packaging marine microorganisms according to the disclosed method allows for widespread distribution to consumers through conventional retail sales channels.

Abstract: The present invention relates to the preparation of universal inactivated vaccines and their use in preparing compositions for the prophylaxis and therapy of dermatomycosis. Vaccines according to the present invention have the advantage of conferring immunity against all important causes of dermatomycosis in animals and are characterized by stable immunogenic properties, easy preparation, high content of microconidia and lack of side reactions in animals.

Abstract: Disclosed is a biopharmaceutical product cryopreservation system, for cryopreserving a biopharmaceutical product that includes a cryopreservation compartment; a cryopreservation fluid located within the cryopreservation compartment; and a biopharmaceutical product cryopreservation vial located within the cryopreservation compartment and surrounded by the cryopreservation fluid, and the biopharmaceutical product cryopreservation vial including a body that includes an oblong cross-section defining proximal and distal ends of the body, and at least one nucleating structure, coupled to a distal end of the body, and the body including a cryogenically stable material that is compatible with biopharmaceutical products. Also disclosed are cryopreservation vials and methods of cryopreserving biopharmaceutical products.

Abstract: Biologically-active material can be preserved by a method of desiccation, without lyophilisation, in a matrix of glassy trehalose. The method involves forming a coacervate of the biologically-active material and chitosan and then dehydrating mixture of coacervate and trehalose solution. In a cycle time much shorter than a typical freeze drying process biologically-active material, such as viruses, proteins and nucleic acids, can be preserved to provide a material that can be rehydrated. The invention is especially useful for the production of vaccines from preserved material.

Abstract: A system and method are provided for inoculating a biological reactor having a chamber. The chamber is configured to contain influent and biomass to degrade contaminants in the influent. A vessel is configured to receive influent and biomass from the chamber and to substantially isolate the received influent and biomass from that contained in the chamber. The vessel is also configured to deliver the isolated influent and biomass to the chamber, thereby facilitating inoculation of the biological reactor.

Abstract: Materials which are not themselves storage-stable at room temperature are made suitable for storage by mixing them with a carrier substance and spray drying the resulting mixture so as to form particles containing both the material and the carrier substance in which the carrier substance is in an amorphous, i.e. glassy or rubbery, state. Formation of such a composition greatly enhances stability. The material stored may be a biological material such as an enzyme, the components of a chemical reaction such as reagents for carrying out an assay, or even viable biological cells.

Abstract: A collection container and method for collecting a predetermined volume of a biological sample, and particularly a whole blood sample, includes at least one gene induction blocking agent in an amount effective to stabilize and inhibit gene induction. The gene induction blocking agent is able to stabilize nucleic acids in the biological sample at the point of collection to block ex vivo gene induction in the sample when stored at room temperature. The stabilizing agents include cationic compounds, detergents, particularly cationic detergents, chaotropic salts, ribonuclease inhibitors, chelating agents, organic solvents, organic reducing reagents, and mixtures thereof. The biological sample is collected directly from the animal and immediately mixed with the gene induction blocking agent without any intermediate processing or handling.

Abstract: Methods are provided for cryopreserving plant cells and to methods for recovering viable plant cells from long or short term cryopreservation. Plant cells to be cryopreserved can be grown in culture and pretreated with a solution containing an cryorotective agent and a stabilizer. Pretreated cells are acclimated to a reduced temperature and loaded with a cryoprotective agent such as DMSO, propylene glycol or polyethylene glycol. Loaded cells are incubated with a vitrification solution which, for example, comprises a solution with a high concentration of the cryoprotective agent. Vitrified cells retain less than about 20% water content and can be frozen at cryopreservation temperatures for long periods of time without significantly altering the genotypic or phenotypic character of the cells. Plant cells may also be cryopreserved by lyophilizing cells to a preferable water content of about 40% to about 60% by weight prior to exposure to a vitrification solution or loading agent.

Abstract: A composition for forming a dried lens-shaped pellet and processes therefore, are provided. The composition comprises 20 to 40% by weight albumin, about 2 to 5% by weight starch, about 40 to 90% by weight of sugar and/or salt, and about 102 to 1011 microorganisms per gram of the composition. The microorganisms include bacteria, viruses, yeasts, protozoa, and fungi. A process for producing the pellet includes mixing the microorganisms with the albumin and starch to form a mixture, reducing water activity of the mixture by adding the sugar and/or salt to the mixture, shaping the mixture into pellets and drying the pellets under vacuum and at a temperature lower than about −10° C. Also a process for producing a suspension of microorganisms includes resuspending the pellet into a suitable medium. Further, other processes using the pellet include fermentation, reconstituting for purposes of testing, and preserving.

Abstract: With a method for cryo-preservation, at least one specimen is arranged on a storage substrate and specimen data, which are characteristic for features of the specimen, are stored at specific positions. Also, a storage substrate for cryo-preservation with such a method is described.

Abstract: A transport/preservation system has been found which provides safe, effective transport for patient specimens and bodily fluids, and functions as a preservative for biological reagents, therapeutics and personal care products. The transport/preservative formulations can also be used as safe, effective sanitizers on surfaces, equipment, and appliances. The transport preservation formulations comprising a biguanide and one or more other antimicrobial agents are cidal to microorganisms when present in a sample or on a surface to be sanitized.

Abstract: Immobilized cell beads incorporating formulated microbial consortium comprising a synergistic mixture of the following bacterial strains namely, Enterobacter sakazaki, Pseudomonas aeruginosa and Aeromonas sobria selected from the following isolated bacterial strains namely, Yersinia enterocolitica, Aeromonas sobria, Klebsiella pneumoniae, Serratia liquefaciens, Enterobacter sakazaki, Citrobacter amalonaticus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Enterobacter cloaca, Acinetobacter calcoaceticus are prepared, the formulated microbial consortium is immobilized in an appropriate immobilizing agent resulting in the formation of beads and the beads are used for instant BOD estimation using an electronic device and the formulated cell beads are reusable and are capable of assimilating most of the organic matter present in varied industrial effluents.

Abstract: The present invention addresses the need to improve the long-term storage stability (i.e. infectivity) of vector formulations. In particular, it has been demonstrated that for adenovirus, the use of bulking agents, cryoprotectants and lyoprotectants imparts desired properties that allow both lyophilized and liquid adenovirus formulations to be stored at 4° C. for up to 6 months and retain an infectivity between 60-100% of the starting infectivity.

Abstract: A method for preserving immobilized or unimmobilized microbial cells having nitrilase activity and for stabilizing the nitrilase activity of unimmobilized or immobilized microbial cells has been developed. Aqueous suspensions containing at least 100 mM bicarbonate, carbonate, or carbamate salts limit microbial contamination of the stored enzyme catalyst, as well as stabilize the desired nitrilase activity of the unimmobilized or immobilized cells. Microorganisms which are characterized by an nitrilase activity and are stabilized and preserved by this method include Acidovorax facilis 72-PF-15 (ATCC 55747), Acidovorax facilis 72-PF-17 (ATCC 55745), Acidovorax facilis 72W (ATCC 55746), and transformed microbial cells having nitrilase activity, the host cells transformed with Acidovorax facilis 72W nitrilase activity. Especially preferred is an embodiment using ammonium carbamate as the inorganic salt.

Abstract: A serum-free insect cell culture medium is provided which provides improvements in the maximum insect cell density and replication of insect viruses within these cells, at a significantly lower cost than commercially-available media. A method is provided for preparing a lipid emulsion for use in insect culture media for large scale culture, and which produces a more stable emulsion with a longer shelf life. The method is preferably carried out by combining in an organic solvent a surfactant and a mixture of lipids to make a lipid phase, heating the lipid phase to a temperature of 40 to 70 degrees Celsius to form an anhydrous lipid phase and adding an aqueous phase thereto to provide a stable lipid micro-emulsion.

Abstract: A biologically-active material comprising a live virus or mycoplasma is preserved by a method of desiccation, without lyophilisation, in a matrix of glassy trehalose having a residual moisture content of not greater than 2%. The method comprises two vacuum drying stages. In a cycle time much shorter than a typical freeze drying process a virus or mycoplasma can be preserved to provide a material that can be rehydrated to give a vaccine having potency.

Abstract: Gel-based medium compositions and a method of use thereof in normothermic, hypothermic or cryopreservative storage and transport of cell samples are described. These gel-based compositions contain a cell maintenance and preservation medium together with a gelling agent. Such gel-based medium compositions protect various cell samples, such as animal or plant organs, tissues and cells, from the mechanical, physiological and biochemical stresses inherently associated with liquid preservation techniques.

Abstract: The invention relates to a process for a simplified and rapid preparation of sterile culture media using microwave ovens. The sterile media of the invention are usable for the selection and identification of microorganisms transformed by viral or plasmid DNA.

Abstract: Linear polymers of glycerol can prevent or delay ice nucleation in a variety of contexts. Polyglycerol can also be employed in combination with other ice control agents, such as polyvinyl alcohol/polyvinyl acetate copolymers and antifreeze proteins, to provide antinucleation effects that are superior to those of either polyglycerol or the co-antinucleator alone. Polyglycerol has a number of advantageous physical and toxicological properties, such as extreme water solubility, non-toxicity to human beings, non-toxicity to animal tissues and organs in vitro even at extreme concentrations, minimal foaming tendency, minimal retention on hydrophobic surfaces, and stability in solution without the need for periodic heating to reactivate its antinucleation properties.

Abstract: A collection container and method for collecting a predetermined volume of a biological sample, and particularly a whole blood sample, includes at least one gene induction blocking agent in an amount effective to stabilize and inhibit gene induction. The gene induction blocking agent is able to stabilize nucleic acids in the biological sample at the point of collection to block ex vivo gene induction in the sample when stored at room temperature. The stabilizing agents include cationic compounds, detergents, particularly cationic detergents, chaotropic salts, ribonuclease inhibitors, chelating agents, organic solvents, organic reducing reagents, and mixtures thereof. The biological sample is collected directly from the animal and immediately mixed with the gene induction blocking agent without any intermediate processing or handling.

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: A collection container and method for collecting a predetermined volume of a biological sample, and particularly a whole blood sample, includes an effective amount of at least one stabilizing agent. The stabilizing agent is able to stabilize nucleic acids in the biological sample at the point of collection to prevent enzymatic degradation of the nucleic acids. The stabilizing agents include cationic compounds, detergents, particularly cationic detergents, chaotropic salts, ribonuclease inhibitors, chelating agents, and mixtures thereof.

Abstract: Compositions and methods for the reversible preservation of biological samples are provided. The compositions include Acacia Gum, including derivations and modifications thereof which are useful as a reversible preservation solution. A method is provided for using Acacia Gum to isolate and reversibly preserve a biological specimen in a dormant state at room temperature for an extended period with minimal damage to the specimen. The compositions and methods disclosed may also be used to create reversibly preserved biological specimens and biological receptors for use in biosensors.

Abstract: A two stage method of thawing cells from a cryopreserved state includes first warming the cells from a cryopreservation temperature to a transition temperature of at least −30° C. in a first, slow-warming stage by exposing the cells to an atmosphere having a temperature of less than 30° C., and once the cells have obtained the transition temperature, subsequently further warming the cells from the transition temperature by exposing the cells to a temperature of at least 32° C. in a second, rapid-warming stage. After the cells obtain the transition temperature in the first stage, the cells may be equilibrated at the transition temperature for a period of time prior to conducting the second stage warming. The method is particularly useful in warming cryopreserved cells attached to a fixed substrate. A thermal conduction device in association with the cryopreserved cells may also be used to further assist in the warming procedure.

Abstract: A process for the preparation of L-amino acids, in which the following steps are carried out, a) fermenting the desired L-amino acid-producing bacteria in which at least the glyA gene is attenuated, in particular by removal of the natural promoter, and optionally b) concentrating the desired product in the medium or in the cells of the bacteria and c) isolating the L-amino acid, and optionally bacteria in which further genes of the biosynthesis pathway of the desired L-amino acid are additionally amplified are employed, or bacteria in which the metabolic pathways which reduce the formation of the desired L-amino acid are at least partly eliminated are employed, and nucleotide sequences of the lacI-tac-5′glyA or lacI-tac-glyA unit.

Abstract: Methods and compositions for cryopreserving somatic cells are provided. In one embodiment, a cell cryopreservation medium is provided which includes an effective amount of arabinogalactan to maintain the viability of cells upon freezing, storage and thawing. The cells may be cooled or frozen during storage to a temperature about or below 4° C., for example, to about −196° C. In one preferred embodimnent, ultrarefined arabinogalactan is provided in the cryopreservation medium, optionally in combination with a second cryopreservation agent, such as dimethyl sulfoxide. The medium can be used for the cryopreservation of a wide variety of different cell types from different sources. For example, mammalian cells, including porcine, canine, human, equine, rodent and bovine cells can be cryopreserved in the medium. The presence of arabinogalactan in the medium protects the viability of cells in the medium during the process of freezing, storage and thawing.

Abstract: A method for arousing dormant bacteria. The method comprises inducing diffusion of intracellular solutes from dormant bacteria and then allowing an adjustment period for a length of time sufficient to initiate arousal. The decrease in intracellular osmolality or pH can be induced by methods such as extraction, dilution, or dialysis. The method has been standardized using Dulbecco's phosphate buffered saline as the solution. The aroused bacteria can then be selected or recovered by growing them on media for a period of time. If the adjustment period is prolonged, many bacteria can become hypermutative.

Abstract: A method of preparing biological materials for cryopreservation is presented. The method lessens the amount of heat released by a cryoprotectant during a latent heat phase by freezing the protectant, thawing the protectant, and treating biologically active materials with the thawed protectant. First, the protectant is frozen to induce an irreversible phase change, along with an irreversible release of energy. After this phase change has occurred, the protectant is thawed and used to treat viable cells or other biologically active material about to undergo freezing. The thawed protectant within the biologically active cells has a reduced endothermic reaction upon subsequent freezing.

Abstract: The present invention discloses: (i) a non-pathogenic probiotic microorganism and its probiotic/therapeutic uses; (ii) a formulation comprising an aqueous solution of a volatile fraction (VF) prepared from the extract of at least one plant derived material and its therapeutic uses; (iii) a process of manufacturing the formulation from the plant derived material; (iv) a probiotic composition comprising the non-pathogenic probiotic microorganism of the invention and/or other probiotic microorganism(s) and the formulation of the invention, and its probiotic/therapeutic uses; (v) a composition for industrial applications comprising the formulation of the invention and microorganism(s) of industrial applicability; and (vi) industrial processes and apparatuses in which the latter composition is used.

Abstract: A new isolate of Ehrlichia species has been obtained from a patient suffering from ehrlichiosis. The new isolate has been found to be similar, but distinctly different from E. canis. A diagnostic kit and methods for diagnosing ehrlichiosis in humans and for screening drugs toxic to the new isolate have been described.

Abstract: The present invention discloses: (i) a non-pathogenic probiotic microorganism and its probiotic/therapeutic uses; (ii) a formulation comprising an aqueous solution of a volatile fraction (VF) prepared from the extract of at least one plant derived material and its therapeutic uses; (iii) a process of manufacturing the formulation from the plant derived material; (iv) a probiotic composition comprising the non-pathogenic probiotic microorganism of the invention and/or other probiotic microorganism(s) and the formulation of the invention, and its probiotic/therapeutic uses; (v) a composition for industrial applications comprising the formulation of the invention and microorganism(s) of industrial applicability; and (vi) industrial processes and apparatuses in which the latter composition is used.

Abstract: Live bacteria are preserved by subjecting drying an aqueous system containing the growing bacteria at room temperature, without special equipment, in the presence of trehalose with or without the addition of divalent cations as stabilizing agents.

Abstract: A new isolate of Ehrlichia species has been obtained from a patient suffering from ehrlichiosis. The new isolate has been found to be similar, but distinctly different from E. canis. A diagnostic kit and methods for diagnosing ehrlichiosis in humans and for screening drugs toxic to the new isolate have been described.

Abstract: The present invention discloses: (i) a non-pathogenic probiotic microorganism and its probiotic/therapeutic uses; (ii) a formulation comprising an aqueous solution of a volatile fraction (VF) prepared from the extract of at least one plant derived material and its therapeutic uses; (iii) a process of manufacturing the formulation from the plant derived material; (iv) a probiotic composition comprising the non-pathogenic probiotic microorganism of the invention and/or other probiotic microorganism(s) and the formulation of the invention, and its probiotic/therapeutic uses; (v) a composition for industrial applications comprising the formulation of the invention and microorganism(s) of industrial applicability; and (vi) industrial processes and apparatuses in which the latter composition is used.

Abstract: The present invention discloses: (i) a non-pathogenic probiotic microorganism and its probiotic/therapeutic uses; (ii) a formulation comprising an aqueous solution of a volatile fraction (VF) prepared from the extract of at least one plant derived material and its therapeutic uses; (iii) a process of manufacturing the formulation from the plant derived material; (iv) a probiotic composition comprising the non-pathogenic probiotic microorganism of the invention and/or other probiotic microorganism(s) and the formulation of the invention, and its probiotic/therapeutic uses; (v) a composition for industrial applications comprising the formulation of the invention and microorganism(s) of industrial applicability; and (vi) industrial processes and apparatuses in which the latter composition is used.

Abstract: A method of shelf preserving biologically active specimens by vitrifying them, i.e., dehydrating them in such a way as to achieve a true glass state at storage temperature by subsequent cooling. The method is founded upon the recognition that to store samples in a true glass state the dehydration temperature of the material to be dehydrated must be higher than the suggested storage temperature. Because the vitrification temperature quickly decreases with increasing water content (for example, pure water vitrifies at Tg=−145° C., whereas 80 percent by weight sucrose solution vitrifies at Tg=−40° C. and anhydrous sucrose vitrifies at Tg=60° C.) the sample needs to be strongly dehydrated to increase the Tg above the temperature of storage (Ts). As determined by the inventor, the dehydration temperature should be selected as higher than the suggested storage temperature, and the glass state is subsequently achieved by cooling after dehydration.

Abstract: The present invention is directed to novel substituted &agr;-hydroxy acid thereof, represented by the general Formula I: where R1-R5, X and Z are defined herein. The present invention also relates to the discovery that compounds having Formula I are potent inhibitors of caspases and apoptotic cell death. Therefore, the inhibitors of this invention can retard or block cell death in a variety of clinical conditions in which the loss of cells, tissues or entire organs occurs.

Abstract: A Rhodobacter species and variants thereof are provided for odor remediation of anaerobic livestock waste lagoons. Also provided are vector systems and genetically reconstituted Rhodobacter PS9 cells, and related methods for biomass production in anaerobic livestock waste lagoons.

Abstract: Disclosed is a universally-adaptable cell preconditioning and storage medium that can be used for the effective cryopreservation of cells at temperatures less than 4° C. The aqueous medium contains adenosine, a calcium channel blocker, and a nutrient-rich matrix that has a sufficient amount of cell metabolites to sustain the metabolic needs of the harvested cells while incubating the cells for a period of at least 10 minutes, without producing detectable levels of lactate or substantially depleting the metabolic substrates of the cell.

Abstract: A method of enhancing viability of an oocyte by contacting the oocyte with a Wnt-4 polypeptide.

Abstract: A method and agent for antibodies against Treponema pallidum are provided. The method involves gene-amplifying and cloning a selection of recombinant antigens. The selection of recombinant antigens consists of 17 kD antigen, 47 kD antigen and TmpA. The antigens are expressed in host vector systems, followed by purification. The purified antigens are then bound to a solid phase individually or in combination, and subjected to a reaction with a clinical specimen. The antibodies bound from the clinical specimen by means of an antigen/antibody reaction are determined by means of a detection system wherein the selection of the recombinant antigens for detecting antibodies to Treponema pallidum consists of 17 kD antigen, 47 kD antigen and TmpA.

Abstract: This invention provides methods of drying and stabilizing prokaryotic cells, and the compositions obtained thereby. The cells are first cultured or incubated under conditions sufficient to induce intracellular trehalose, suspended in a stabilizing solution and dried to form a solid glass. The resulting product is storage-stable at room temperature, showing little viability loss on storage.

Abstract: The present invention provides a method for carrying out in vitro the complete developmental sequence culture of tissular parasites, which includes culturing the parasites in a totally defined culture medium which is an axenic monophasic liquid culture medium, free of serum and free of serum-derived or cell-derived macromolecules, proteins and peptides. For obtaining amastigote forms, this medium is buffered at a pH of 5.5 to 6.5 and has an osmolarity of at least 400 milliosmoles/kg of liquid. For obtaining promastigote forms, the medium is buffered at a pH of 7 to 7.5 and has an osmolarity of at least 300 milliosmoles/kg liquid. Application to the in vitro culture of different stages of tissular parasites such as Leishmania, T. cruzi, and hamatoprotozoa is also provided.

Abstract: Disclosed is a biopharmaceutical product cryopreservation system, for cryopreserving a biopharmaceutical product that includes a cryopreservation compartment; a cryopreservation fluid located within the cryopreservation compartment; and a biopharmaceutical product cryopreservation vial located within the cryopreservation compartment and surrounded by the cryopreservation fluid, and the biopharmaceutical product cryopreservation vial including a body that includes an oblong cross-section defining proximal and distal ends of the body, and at least one nucleating structure, coupled to a distal end of the body, and the body including a cryogenically stable material that is compatible with biopharmaceutical products. Also disclosed are cryopreservation vials and methods of cryopreserving biopharmaceutical products.

Abstract: An improved preservation solution is described, which is intended for the preservation of organs and tissues, or parts thereof, from humans and animals. The improved preservation solution contains calcium, at least one colloidosmotically active substance, and nitroglycerin. Also described is a method for preserving organs and tissues, or parts thereof, from humans and animals in the improved preservation solution.

Abstract: Materials which are not themselves storage-stable at room temperature are made suitable for storage by mixing them with a carrier substance and spray drying the resulting mixture so as to form particles containing both the material and the carrier substance in which the carrier substance is in an amorphous, i.e. glassy or rubbery, state. Formation of such a composition greatly enhances stability. The material stored may be a biological material such as an enzyme, the components of a chemical reaction such as reagents for carrying out an assay, or even viable biological cells.

Abstract: Third stage juvenile (J3) entomopathogenic nematodes are prepared for storage by being induced into a state of cryptobiosis. The induction of cryptobiosis is effected by mixing an aqueous cream of the J3 nematodes with anhydrous, small particles (average maximum dimension less than 300 &mgr;m) of non-fibrous cellulose. The proportions of the aqueous cream and non-fibrous cellulose particles are such that, after equilibration, the mixture has a water activity in the range 0.80 to 0.995. Preferably an anti-fungal agent is included in the aqueous cream. To store the cryptobiotic J3 nematodes, the mixture is preferably kept in a container, fitted with an attachment which maintains the water activity in the container at a required value. The attachment includes a rigid tube that connects the interior of the container with a chamber that is vented to ambient atmosphere by small apparatus.