Abstract: Improved cloning vectors derived from pRO1614 are described. One of these vectors, pRO1727, is suitable for cloning using DNA cleaved with the restriction endonuclease, PstI, and allows selection for the recovery of recombinant plasmids using tetracycline resistance. The cloning efficiency observed for pRO1727 is higher than described previously for pRO1614 and the host range of this vector is now restricted to Pseudomonas bacteria. Another vector, designated pRO1729, is described and developed from pRO1727 by deletion of a portion of its DNA and incorporation of a segment of DNA which encodes for resistance to the antibiotic, chloramphenicol. The chloramphenicol resistance determinant has a cleavage site for restriction endonuclease EcoRI within its chloramphenicol resistance determinant. Thus, DNA cloned into this site results in the loss of chloramphenicol resistance which can be detected subsequent to a cloning experiment. Both pRO1727 and pRO1729 are more useful in Pseudomonas for cloning than pRO1614.

Abstract: An improved gene splicing and recombinant plasmid transformation method is described. The method includes mechanical fragmenting of chromosomal DNA followed by conventional digestion with a restriction enzyme and gene splicing into a vector to provide recombinant plasmids in a bank of at least about 100 different plasmids. The plasmids in the bank are provided for transformation into a suitable host, particularly a plasmid free bacterium of the same species from which the chromosomal DNA or the vector is derived. The method provides high transformation frequencies because of the presence of multiple "super coiled" or closed coiled recombinant plasmids in the bank. The method also allows for the direct selection of many different phenotypic traits in a pool of the transformed hosts. The selected hosts are useful for the production of various gene products.

Abstract: Non-sucrose fermenting strains of Pediococcus pentosaceus are described. The non-sucrose fermenting Pediococcus pentosaceus strains have been cured to remove a single, natural plasmid which has been found to control the fermentation of sucrose to produce lactic acid. The natural plasmid removed is between about 30 to 35 megadaltons in molecular weight. Compositions including the non-sucrose fermenting strains of Pediococcus pentosaceus are useful for food fermentations, particularly meat fermentations.

Abstract: Plants susceptible to pathogenic fungi are contacted with a mutant strain of Pseudomonas putida, particularly Pseudomonas putida NRRL-B-12537 which produces iron complexing siderophores thereby affording protection from the fungi. The Pseudomonas competes with the fungi for iron found in the soil thereby inhibiting the fungi growth. The method is particularly effective in controlling Fusarium oxysporum Sp lycopersici on tomato plants.

Abstract: A method for using selected strains of Streptococcus lactis subspecies diacetilactis, which have been modified to be non-lactose fermenting, for the preservation of foods containing lactose is described. The subspecies is more generally known as Streptococcus diacetilactis. The selected Streptococcus diacetilactis strains have been modified by curing to remove at least one natural plasmid which controls the fermentation of lactose to lactic acid while retaining the ability of this subspecies to inhibit bacterial spoilage in foods. The plasmid removed by curing is about 41 megadaltons (Mdal) in mass. The method using the modified strains of Streptococcus diacetilactis is particularly adapted for the preservation of milk products.

Abstract: The present invention relates to novel, broad bacterial host range small plasmid deoxyribonucleic acid rings which serve as cloning vehicles for DNA fragments, particularly those separated from other plasmid rings or from chromosones, recombined with the small plasmid rings and to the processes for recombining the plasmid rings and to processes for transferring them between host bacteria. In particular, the present invention relates to the aggregate plasmid ring RP1/pRO1600, to pRO1600 and plasmid ring derivatives thereof, particularly including pRO1601; pRO1613 and pRO1614, all of which are carried for reference purposes in Pseudomonas aeruginosa ATCC 15692 (also known as strain PAO1c) and are on deposit at the Northern Regional Research Laboratories (NRRL) of the United States Department of Agriculture at Peoria, Ill. The plasmid ring RP1 (also known as R1822) is deposited in Pseudomonas aeruginosa NRRL-B-12123 (and is a known plasmid ring). The pRO1600 portion of the aggregate is a new plasmid ring.

Abstract: A method is described for producing fermented foods by generating lactic acid in the food using a culture of a lactobacillus similar to Lactobacillus casei subspecies alactosus NRRL-B-12,344 and a stimulatory food grade metal salt, wherein the culture has unique rapid low temperature fermentation characteristics and wherein lactose, glycogen, and starch are not fermented by the culture. The preferred Lactobacillus casei subspecies alactosus is NRRL-B-12,344 or strains having low temperature food fermentation characteristics in common with this strain. In order to provide rapid fermentation, the stimulatory, food grade metal salt, usually a manganese salt, is provided in the food or the culture which is added to the food with the selected lactobacillus to accelerate fermentation. The cultures are particularly suited for the controlled fermentation of carbohydrates, naturally present in or added to the food to provide a selected final pH.

Abstract: A method for the preparation of naturally stabilized, thick bodied, fermented milk products by fermentation is described. Mixed cultures of milk fermenting, non-slime, lactic acid producing bacteria and slime producing Streptococcus lactis, Streptococcus cremoris or mixtures thereof having the thickening characteristics in milk of Streptococcus cremoris NRRL-B-12,361, 12,362 or 12,363 are used, preferably in addition with a diacetyl producing bacterium for flavor. The fermented milk products are thick bodied without any ropiness or sliminess and are stable to separation of whey from curd upon storage at refrigeration temperatures, with little or no added stabilizing agents such as gums and starches or thickening agents such as added non-fat milk solids. The preferred product is a thick bodied buttermilk.

Abstract: The present invention relates to novel, broad bacterial host range small plasmid deoxyribonucleic acid rings which serve as cloning vehicles for DNA fragments, particularly those separated from other plasmid rings or from chromosones, recombined with the small plasmid rings and to the processes for recombining the plasmid rings and to processes for transferring them between host bacteria. In particular, the present invention relates to the aggregate plasmid ring RP1/pRO1600, to pRO1600 and plasmid ring derivatives thereof, particularly including pRO1601; pRO1613 and pRO1614, all of which are carried for reference purposes in Pseudomonas aeruginosa ATCC 15692 (also known as strain PAO1c) and are on deposit at the Northern Regional Research Laboratories (NRRL) of the U.S. Department of Agriculture at Peoria, Il. The plasmid ring RP1 (also known as R1822) is deposited in Pseudomonas aeruginosa NRRL-B-12123 (and is a known plasmid ring). The pRO1600 portion of the aggregate is a new plasmid ring.

Abstract: The present invention relates to novel, broad bacterial host range small plasmid deoxyribonucleic acid rings which serve as cloning vehicles for DNA fragments, particularly those separated from other plasmid rings or from chromosones, recombined with the small plasmid rings and to the processes for recombining the plasmid rings and to processes for transferring them between host bacteria. In particular, the present invention relates to the aggregate plasmid ring RP1/pRO1600, to pRO1600 and plasmid ring derivatives thereof, particularly including pRO1601; pRO1613 and pRO1614, all of which are carried for reference purposes in Pseudomonas aeruginosa ATCC 15692 (also known as strain PAO1c) and are on deposit at the Northern Regional Research Laboratories (NRRL) of the U. S. Dept. of Agriculture at Peoria, Ill. The plasmid ring RP1 (also known as R1822) is deposited in Pseudomonas aeruginosa NRRL-B-12123 (and is a known plasmid ring). The pRO1600 portion of the aggregate is a new plasmid ring.

Abstract: A method is described for producing fermented vegetables by generating lactic acid in an acidic brine solution using selected cultures of Pediococcus pentosaceus which have rapid low temperature fermentation characteristics. The preferred Pediococcus pentosaceus is NRRL-B-11,465 which effectively and rapidly removed brine carbohydrate thus lowering the pH at unusually low brine temperatures less than about 25.degree. C. (77.degree. F.) and at high salinity. A stimulatory food grade, metal salt, preferably a manganese salt, is provided in the acidic brine solution with the Pediococcus pentosaceus to accelerate growth and reduce the fermentation time. The method is particularly adapted to the controlled fermentation of cucumbers in making pickles.

Abstract: Naturally stabilized fermented milk products are prepared with a concentrate of Streptococcus thermophilus cells that produce a stabilizer in situ when cultured in milk. The concentrate is obtained by culturing the stabilizer-producing Streptococcus thermophilus in a growth medium including milk solids to obtain at least about 10.sup.8 cells per ml. The growth medium preferably contains maltose, sucrose, fructose or lactose which enhances stabilizer formation.

Abstract: An improved process for producing a superior flavor in fermented meat, particularly sausage, using Micrococcus varians in admixture with Pediococcus cerevisiae NRRL-B-5627 and/or with other lactic acid producing meat fermenting bacteria for lowering the pH is described. Micrococcus varians is a very poor producer of lactic acid and alone cannot produce acceptable sausage. The improved bacterial compositions develop the solid bright red color associated with sausage and other fermented meats in the presence of edible nitrate and/or edible nitrite. Combinations of the nitrite and nitrate can be used. No acid forming chemicals, such as gluconic acid delta lactone, are used in the fermentation process. The bacterial composition is preferably provided in the form of a frozen cell concentrate for storage and subsequent thawing for use.

Abstract: A method and bacterial compositions are described for producing fermented meat by generating lactic acid using selected cultures of Pediococcus pentosaceus which have unique low temperature meat fermentation characteristics. The preferred Pediococcus pentosaceus is NRRL-B-11,465 which is unusually rapid in lowering the pH at low meat temperatures. A stimulatory, edible metal salt, preferably a manganese salt, is provided in the meat with the Pediococcus pentosaceus to reduce or eliminate preservative inhibition and/or to accelerate growth at meat temperatures between 15.6.degree. C. (60.degree. F.) to 26.7.degree. C. (80.degree. F.). Preservatives to prevent spoilage and rancidity can be used in the meat and can include hydroxyaryl antioxidants, particularly butylated hydroxytoluene (BHT) and/or butylated hydroxyanisole (BHA). BHT and/or BHA together with other meat preservatives, particularly sodium chloride, severely inhibit NRRL-B-11,465 at low meat temperatures between 15.6.degree. C. (60.degree. F.

Abstract: A food for carnivorous lower animals which is fermented by certain lactic acid producing bacteria that produce a desirable flavor or which has an added flavoring from an edible material fermentate of lactic acid producing bacteria is described. The pet food has a pH of from about 3 to less than 6 and is particularly palatable to dogs.

Abstract: Storage stable, lyophilized, acid producing bacteria, such as lactic acid bacteria, are prepared by lyophilizing the bacteria in the presence of at least 5% by weight of the bacteria of a basic organic or inorganic buffering agent, preferably an alkali metal salt of glycerophosphate, to provide lyophilized bacteria containing less than 5% by weight water, and sealing the lyophilized bacteria in a container while in a vacuum or surrounded by a gaseous atmosphere substantially free of oxygen, preferably argon which is essentially free of oxygen.

Abstract: A biologically concentrated Streptococcus diacetylactis is used to develop in creamed cottage cheese a desirable diacetyl flavor and aroma when creamed cottage cheese is stored at refrigeration temperatures. The biologically concentrated Streptococcus diacetylactis is prepared without mechanical concentration by growing Streptococcus diacetylactis on a special media to produce at least about 10.sup.9 cells per ml.

Abstract: A novel mutant strain Streptococcus diacetilactis NRRL-B-8177 which is particularly adapted for the preparation of creamed Cottage cheese without fermentation (or prior incubation) of the creaming mixture or dressing is described. This strain produces about one-half of the acid produced by S. diacetilactis 18-16 which is regarded as the best bacterium that is commercially available for use in Cottage cheese. Improved flavor and prophylaxis against spoilage bacteria of the dressed Cottage cheese is preferably provided by blending a concentrate of the Streptococcus diacetilactis NRRL-B-8177 cells with a creaming mixture at less than about 50.degree. F. (10.degree. C.) and then mixing the cold cream mixture with dry Cottage cheese curd cooled to less than about 50.degree. F. and maintaining the temperature at less than about 50.degree. F.

Abstract: A food for carnivorous lower animals which is fermented by certain lactic acid producing bacteria that produce a desirable flavor or which has an added flavoring from an edible material fermentate of lactic acid producing bacteria is described. The pet food has a pH of from about 3 to less than 6 and is particularly palatable to dogs.

Abstract: An improved process for producing a superior flavor in fermented meat, particularly sausage, using Micrococcus varians in admixture with Pediococcus cerevisiae NRRL-B-5627 and/or with other lactic acid producing meat fermenting bacteria for lowering the pH is described. Micrococcus varians is a very poor producer of lactic acid and alone cannot produce acceptable sausage. The improved bacterial compositions develop the solid bright red color associated with sausage and other fermented meats in the presence of edible nitrate and/or edible nitrite. Combinations of the nitrite and nitrate can be used. No acid forming chemicals, such as gluconic acid delta lactone, are used in the fermentation process. The bacterial composition is preferably provided in the form of a frozen cell concentrate for storage and subsequent thawing for use.