CO2-mediated bloater defect can be induced by the uncontrolled growth of Enterobacteriaceae in cucumber fermentation.

Enterobacteriaceae are known to proliferate in cucumber juice, deriving energy from the fermentation of sugars to organic acids and ethanol, and theoretically generating carbon dioxide (CO2). We hypothesized that the CO2 produced by the indigenous Enterobacteriaceae in the early stage of cucumber fermentation accumulates in the fermenting fruits causing bloater defect. The ability of seven Enterobacteriaceae, indigenous to cucumber, to grow and produce CO2 in cucumber juice medium (CJM), a sterile model system for cucumber fermentation, was characterized. The induction of bloater defect in cucumber fermentation conducted with pasteurized and acidified fruits was also evaluated. The generation times of the seven Enterobacteriaceae in CJM ranged between 0.25 and 8.20 h and resulted in carbon dioxide (CO2) production to estimated amounts of 7.22-171.5 mM. Enterobacter cancerogenus and Enterobacter nimipressuralis were among the bacteria that produced the most and the least CO2 in CJM, respectively, at estimated mM concentrations of 171.58 ± 42.96 and 16.85 ± 6.53. Inoculation of E. cancerogenus and E. nimipressuralis in acidified and pasteurized cucumbers resulted in the production of 138 and 27 mM CO2, respectively. Such Enterobacteriaceae produced 2% hydrogen in the model cucumber fermentations. A bloater index of 25.4 and 17.4 was calculated from the cucumbers fermented by E. cancerogenus and E. nimipressuralis, respectively, whereas no defect was observed in the fruits collected from uninoculated control fermentation jars. It is concluded that the metabolic activity of the Enterobacteriaceae indigenous to cucumber can produce sufficient CO2 in cucumber fermentations to induce bloater defect.

Growth of ɣ-Proteobacteria in Low Salt Cucumber Fermentation Is Prevented by Lactobacilli and the Cover Brine Ingredients.

This study investigated the ability of É£-proteobacteria, indigenous to fresh cucumber, to grow in the expressed fruit juice (CJM) and fermentation. It was hypothesized that fresh cucumbers can support prolific growth of É£-proteobacteria but that the cover brine composition and acid production by the competing lactobacilli in the fermentation of the fruit act as inhibitory agents. The É£-proteobacteria proliferated in CJM with an average maximum growth rate (µmax) of 0.3895 ± 0.0929 and doubling time (Td) of 1.885 ± 0.465/h. A significant difference was found between the É£-proteobacteria µmax and Td relative to Lactiplantibacillus pentosus LA0445 (0.2319 ± 0.019; 2.89/h) and Levilactobacillus brevis 7.2.43 (0.221 ± 0.015; 3.35/h) but not Lactiplantibacillus plantarum 3.2.8 (0.412 ± 0.119; 1.87/h). While inoculation level insignificantly altered the µmax and Td of the bacteria tested; it impacted the length of lag and stationary phases for the lactobacilli. Unlike the lactobacilli, the É£-proteobacteria were inhibited in CJM supplemented with a low salt fermentation cover brine containing calcium chloride, acetic acid and potassium sorbate. The É£-proteobacteria, P. agglomerans, was unable to proliferate in cucumber fermentations brined with calcium chloride at a pH of 6.0 ± 0.1 and the population of Enterobacteriaceae was outcompeted by the lactobacilli within 36 h. Together these observations demonstrate that the prolific growth of É£-proteobacteria in CJM is not replicated in cucumber fermentation. While the É£-proteobacteria growth rate is faster that most lactobacilli in CJM, their growth in cucumber fermentation is prevented by the cover brine and the acid produced by the indigenous lactobacilli. Thus, the lactobacilli indigenous to cucumber and cover brine composition influence the safety and quality of fermented cucumbers. IMPORTANCE While the abundance of specific É£-proteobacteria species varies among vegetable type, several harbor Enterobacteriaceae and Pseudomonadaceae that benefit the plant system. It is documented that such bacterial populations decrease in density early in vegetable fermentations. Consequently, it is assumed that they do not contribute to the quality of finished products. This study explored the viability of É£-proteobacteria in CJM, used as a model system, CJM supplemented with fermentation cover brine and cucumber fermentation, which are characterized by an extremely acidic endpoint pH (3.23 ± 0.17; n = 391). The data presented demonstrates that fresh cucumbers provide the nutrients needed by É£-proteobacteria to proliferate and reduce pH to 4.47 ± 0.12. However, É£-proteobacteria are unable to proliferate in cucumber fermentation. Control of É£-proteobacteria in fermentations depends on the cover brine constituents and the indigenous competing lactobacilli. This knowledge is of importance when developing guidelines for the safe fermentation of vegetables, particularly with low salt.

Vegetable fermentations brined with low salt for reclaiming food waste.

Fermentation of eight vegetables was studied as an alternative for reclamation of surplus volumes. Fermentation performance was predicted by comparing the amounts of acid that could be produced from the intrinsic sugar content with that buffered by the fresh vegetable matrices prior to reaching an inhibitory pH for fermentative microbes (3.30). Native fermentations were brined with 345.0 mM sodium chloride, 40.0 mM calcium chloride, 6.0 mM potassium sorbate, and vinegar to adjust the initial pH to 4.70. High-performance liquid chromatography analysis, pH, and carbon dioxide measurements and spiral plating on selective media were employed to monitor the progress of fermentations. The average colony counts for yeast and/or molds and Enterobacteriaceae declined to undetectable levels from 3.6 ± 1.5 log CFU/ml within 7 days of fermentation. The fermentation of sugars produced lactic, acetic, succinic, and/or malic acids, and ethanol. As predicted, the fermentation of vegetables with low sugar content, such as broccoli, green leaf lettuce, and green pea proceeded to completion. The fermentation of vegetables with a moderate sugar content, such as green bell pepper, red ripened tomato, and green bean were incomplete at pH 3.1 ± 0.2. The fermentation of high sugar vegetables including sweet potato and corn were expected and observed to be incomplete. It is concluded that the intrinsic sugar content and buffer capacity of surplus vegetables are relevant parameters in obtaining complete fermentations. PRACTICAL APPLICATION: Vegetables are the second most wasted commodity in the United States and a substantial constituent of the global food waste. Development of fermentation to reclaim surplus vegetables from farms, grocery stores, and farmer's markets offers opportunities to ameliorate economic losses and environmental impact and add value to waste. The research described here suggests that a fraction of vegetables could be fermented in cover brines while others, with high sugar content, need specialized handling. Evidently, optimization of vegetable fermentation with starter cultures and added buffers represent an opportunity to stimulate complete bioconversions useful for reclaiming surplus volumes.

Genotypic and phenotypic diversity among Lactobacillus plantarum and Lactobacillus pentosus isolated from industrial scale cucumber fermentations.

The Lactobacillus plantarum and Lactobacillus pentosus genotypes existing in industrial-scale cucumber fermentations were defined using rep-PCR-(GTG)5. The ability of each genotype to ferment cucumbers under various conditions was evaluated. Rep-PCR-(GTG)5 was the technique capable of illustrating the most intraspecies discrimination compared to the sequencing of housekeeping genes (recA, dnaK, pheS and rpoA), MLST and RAPD with primers LP1, OPL5, M14 and COC. Ten genotypic clusters were defined for the 199 L. pentosus tested and three for the 17 L. plantarum clones. The ability of the 216 clones genotyped and 37 additional cucumber fermentation isolates, of the same species, to rapidly decrease the pH of cucumber juice medium under various combinations of sodium chloride (0 or 6%), initial pH (4.0 or 5.2) and temperatures (15 or 30 °C) was determined using a fractional factorial screening design. A reduced fermentation ability was observed for the L. plantarum strains as compared to L. pentosus, except for clone 3.2.8, which had a ropy phenotype and aligned to genotypic cluster A. L. pentosus strains belonging to three genotypic clusters (B, D and J) were more efficient in cucumber juice fermentation as compared to most L. plantarum strains. This research identified three genetically diverse L. pentosus strains and one L. plantarum as candidates for starter cultures for commercial cucumber fermentations.