The ecology of the streptococci.

All species of streptococci are divided into several groups according to the ecological niches they occupy. Members of the lactic acid group are found only in the plant environment and the dairy. Members of the pyogenic group and the buccal and intestinal residents of the subthermophilic group are found in intimate association only with homeothermic hosts. Members of the virulent pyogenic group, such asStreptococcus pyogenes, S. equi, andS. infrequens, are restricted to single hosts-the human, the horse, and the pig-whereas others, such asS. agalactiae can be found in several hosts. The intestinal dwellers of the subthermophilic group,S. bovis andS. equinus, occur in several animal species and are not isolated from other sources. The superficially homogeneousS. faecalis of the "enteric" group may actually consist of a group specifically restricted to the human host, and of a second group distributed among wild animals, insects, and plants. The latter group differs from the human type by the ability to hydrolyze starch and to produce a rennin-proteolytic type of curd in milk.Streptococcus faecium is widely distributed in both the homeothermic and poikilothermic environments, but no properties which differentiate the sources are known.Streptococcus faecium biot.casseliflavus has been isolated only from insects and plants.Streptococcus uberis occurs on the bovine lips and skin. Unidentified streptococci from all environments vary widely in properties. Many resembleS. lactis, S. faecalis, andS. faecium.

Microbial Changes in Sweet Sorghum (Sorghum bicolor) Juices.

Juice freshly expressed from Sorghum bicolor for making sweet sorghum syrup contained 10 microorganisms per ml. The dominant bacterium was Leuconostoc mesenteroides, followed by gram-negative rods. Lactobacilli, yeasts, and nonfecal coliform bacteria each comprised about 1% of the microbial population. Spoilage of juice, manifested by a sour odor, discoloration, and foaming, occurred between 5 and 12 h at ambient temperatures. Spoilage was correlated with a drop in pH from 4.9 to 4.5 L. mesenteroides was the dominant spoiling agent at 20 degrees C, and Lactobacillus plantarum was the dominant spoiling agent at 32 degrees C, as determined by pure culture studies. Juice may be stored for 14 days at 4 degrees C if promptly refrigerated.

Molds on vegetables at the time of harvest.

The mean numbers of colony-forming units of molds present on vegetables at the time of harvest were in the range of 4.2 X 10(3) to 6.7 X 10(3)/g for all vegetables except cucurbits and asparagus. The numbers were not influenced by cultivars within species, duration of the growing season, distant separation of the growing fields, or elevation above ground. Numbers increased with rainfall during either of 3 days before harvest and decreased when the mean daily temperature exceeded 24 degrees C. The most frequently isolated fungi were Aureobasidium pullulans, Fusarium species, Alternaria tenuis, Epicoccum nigrum, Mucor species, Chaetomium fimeti, Rhizopus nigricans, and Phoma species, in decreasing order. All other molds comprised less than 2% of the isolates. Aspergilli and penicillia occurred sporadically and in low numbers. Fusarium species were dominant on cucurbits.

Bacteria within ovules and seeds.

Surface-sterilized ovules and seeds of 27 species of plants were cultured in the water of syneresis of a nutrient medium low in agar content. Bacteria were obtained from 30% of the ovules, 15% of the seeds of herbaceous plants, 16% of the seeds of woody plants, 5.4% of the overwintered noncereal seeds, and 13.5% of overwintered cereal seeds. In no instance did every ovule or seed of a plant species contain bacteria. No bacteria were obtained from the hard, waxy seeds of mimosa or yellowwood. They were not obtained from ovules with unbroken coats or from seeds with coats that were not ruptured during the swelling of the seed. Only one species of bacteria was recovered in 93% of the instances in which bacteria were obtained. Bacteria were obtained from seeds that were embedded in the acidic parenchyma of the lemon or surrounded by the thickened flesh of the cucurbits. The bacteria were distributed among 19 genera and 46 species. The species isolated in greatest numbers were Bacillus megaterium, B. cereus, Erwinia herbicola, Flavobacterium devorans, and Pseudomonas fluorescens. Bacteria recovered less frequently were in the genera Achromobacter, Acinetobacter, Alcaligenes, Brevibacterium, Corynebacterium, Cytophaga, Leuconostoc, Micrococcus, Nocardia, Proteus, Streptococcus, Streptomyces, and Xanthomonas. Members of 11 genera and 15 species of bacteria were isolated once.

Enterococci in insects.

Enterococci were obtained from 213 of 403 insects cultured during a 14-month period, in numbers from 10(3) to 3 x 10(7)/g of insect. Insects were taken only from nonurban, wild, and cultivated fields and woods. In species of insects carrying them, enterococci were not always present in every individual cultured, and often more than one species of enterococcus occurred within a species. Enterococci were obtained from certain insects taken in the field during the dormant season, suggesting their role as overwintering agents. They were generally present in species feeding on nectar, succulent plant parts, and on and ir forest litter, but not from insects feeding on less succulent leaves and stems. Streptococcus faecalis was recovered from 32%, Streptococcus faecium from 22.4%, and Streptococcus faecium var. casseliflavus from 43.5% of members of the 37 taxa of insects. S. faecalis and S. faecium var. casseliflavus exhibit a high percent of conformity to the properties published for them. The heterogeneity in properties of S. faecium is similar to that found for the species taken from plants. Many fail to grow in broth at 45 C or in broth containing 6.5% NaCl; 50% of the cultures ferment both melezitose and melibiose, and a few ferment neither sugar. The remainder ferment melibiose only. Failure to reduce methylene blue in milk by S. faecalis and S. faecium is correlated with the inability to ferment lactose. More than 93% of the cultures of S. faecalis digest casein in milk from the top downward, following the production of a soft, flowing curd. Because this property is not characteristic of S. faecalis taken from humans, the reaction in litmus milk is suggested as a means of differentiation between cultures of remote and innocent origin in nature and recent, human pollution.

Pediococci residing on plants.

The pediococci residing on plants resemble the lactobacilli, but they differ from the streptococci in their limited distribution and low population level on plants. They are a subgroup within the genus Pediococcus which grow freely in neutral media and require neither NaCl nor CO(2). They are most readily recognized by the ability to initiate growth in liquid media, acidified to pH 5.0, which contain 1.5% sodium acetate. In stained preparations the cells occur singly and in pairs, short chains, and clusters. The occurrence of two-dimensional tetrads may be rare; this varies with the individual culture and with the culture medium. The terminal pH in 2% glucose broth varies from 3.6 to 4.3. Ability to initiate growth at 45 C, production of ammonia from arginine, dissimilation of malate, and fermentation of arabinose are confirmatory characteristics. The subgroup contains only two quite similar, but differentiable, species. P. acidilactici initiates growth at 50 C and produces catalase on heated blood medium but does not produce acid-sensitive catalase; a majority of the strains fail to initiate growth at 10 C and many fail to ferment maltose and lactose. P. pentosaceus initiates growth at 10 C but not at 50 C and produces acid-sensitive catalase; catalase production on heated blood medium is transient; a majority of the cultures ferment maltose, salicin, and trehalose. No carbohydrate serves reliably to differentiate between the species. The guanine plus cytosine ratio of P. pentosaceus deoxyribonucleic acid (DNA) was determined to be 35.1 +/- 1.2 and that of P. acidilactici DNA is 38.5 +/- 0.8.

Lactobacilli on plants.

The distribution, enumeration, and identification of lactobacilli on vegetable plants were studied in an area described geographically as being subtropical and moist. The lactobacilli were obtained, by means of quantitative enrichment procedures in Rogosa's SL broth, from 35.3% of all samples incubated at 32 C, and from 15.4% of the samples incubated at 45 C. Less than 10 lactobacilli/g of plant material were enumerated in 54% of all positive samples. The lactobacilli were found much less frequently and in lower numbers than were streptococci or Leuconostoc mesenteroides. The most frequently isolated lactobacillus was very simular to, but not identical with, Lactobacillus fermenti. It was aerogenic, grew well at both 15 and 45 C, fermented arabinose, lactose, and sucrose, and liberated ammonia from arginine. Of the identified species, L. plantarum, L. fermenti, and L. brevis were the most frequently isolated, whereas L. casei, L. viridescens, L. cellobiosus, L. salivarius, and L. buchneri were obtained from small numbers of samples. The widespread but sporadic distribution of lactobacilli in low numbers seems to indicate that these organisms do not normally thrive on plant surfaces. A ternary cycle, beginning with intestinal waste, followed by mechanical distribution to and among plants, and return to the host via the oral cavity, is suggested.

Streptococcus faecium var. casselifavus, nov. var.

Streptococcus faecium var. casseliflavus is a gram-positive, spherical cell. The cells occur chiefly as pairs within chains and elongate to ogive-shaped cells during growth. Growth is good on 5% bile salts-agar and in broth at 10 C, and in broth adjusted to pH 9.6 or containing 6.5% NaCl, but many strains fail to grow at 45 C. Litmus is reduced rapidly prior to formation of an acid curd. Few strains release ammonia from arginine or serine. The organism is not proteolytic and does not produce H(2)S or acetylmethylcarbinol, reduce nitrate, decarboxylate tyrosine, or produce slime on sucrose-agar. Most strains survive heating to 60 C for 30 min. It produces gray colonies on potassium tellurite agar, reduces 2,3,5-triphenyltetrazolium-HCl to a pink color, and ferments cellobiose, dextrin, maltose, mannose, and sorbitol, thus resembling S. faecalis. Like S. faecium, it produces peroxidase but not catalase on heated blood media, dissimilates malate, and ferments arabinose, melibiose, and salicin, but not melezitose. Like both species, it ferments dextrose, galactose, lactose, mannitol, sucrose, trehalose, and citrate. Properties peculiar to the variant include the high pH limiting initiation and termination of growth; the fermentation of alpha-methyl-d-glucoside, raffinose, and xylose; motility; and growth without blue button formation in ethyl violet broth. The water-soluble, pale lemon-yellow pigment is released into the aqueous phase only after the cell envelope is altered by fat solvents. The bacterium thrives as an epiphyte on plants.

Spherical Lactic Acid-producing Bacteria of Southern-grown Raw and Processed Vegetables.

The frequency and levels of population of the spherical lactic acid-producing bacteria were determined on raw and processed yellow summer and zucchini squash, a variety of greens, green beans, okra, southern peas, and butter and lima beans, and on fresh cucumbers and corn flowers. Six taxa occurred consistently: Leuconostoc mesenteroides, yellow-pigmented streptococci, Streptococcus faecium, Aerococcus viridans, and S. faecalis and S. faecalis var. liquefaciens. The same taxa occurred with the same order of frequency on processed, frozen vegetables, but with a marked decrease in the occurrence of S. faecalis var. liquefaciens. S. lactis, S. cremoris, S. equinus, S. bovis, and pediococci were isolated infrequently. No other member of the viridans group of the streptococci and no member of the pyogenic group was isolated. Approximately 88% of the cultures were identified. Total counts of the lactic-acid-producing bacteria rarely exceeded 10 per gram of sample, and there was a reduction by 90% during the second year of study, probably because of drought. Only one bacterial species was found on 40% of the raw and 34% of the processed vegetable samples. Two or more species or taxa were present on the remainder of 153 raw and 56 processed vegetable samples. A. viridans was present on squash, greens, okra, and southern peas, and its frequency of occurrence on vegetables suggests that plants are its natural habitat.

Growth of lactic acid bacteria in waste waters of vegetable-processing plants.

Waters used in washing, blanching, cooling, and conveying vegetables during processing for freezing were filtered, sterilized, and inoculated with Streptococcus faecalis, S. lactis, or Lactobacillus plantarum. The contents of total nitrogen and total solids were determined, and ninhydrin tests and Benedict's tests for reducing sugars were performed. Substances positive to the ninyhydrin tests and also capable of supporting the growth of the bacteria to high levels of population were found in waters used to blanch cut green beans, but not in the cooling or conveying waters. They were found only in waters following slicing of blanched whole beans. They were also present in waters used in processing purple hull peas at all stages, but only in the waters used to blanch and cool lima beans. The substances were present in waters used to wash and blanch squash, but only in the waters used to blanch greens; they were not found in the cooling waters during the handling of either vegetable. No waters used in the processing of okra yielded a positive ninhydrin test, nor did they support the growth of the lactic acid bacteria.