@article{zhai_pagan-medina_perez-diaz_2023, title={CO2-mediated bloater defect can be induced by the uncontrolled growth of Enterobacteriaceae in cucumber fermentation}, ISSN={["2048-7177"]}, DOI={10.1002/fsn3.3557}, abstractNote={Abstract 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.}, journal={FOOD SCIENCE & NUTRITION}, author={Zhai, Yawen and Pagan-Medina, Christian G. and Perez-Diaz, Ilenys M.}, year={2023}, month={Jul} }
 @article{rothwell_zhai_pagan-medina_perez-diaz_2022, title={Growth of gamma-Proteobacteria in Low Salt Cucumber Fermentation Is Prevented by Lactobacilli and the Cover Brine Ingredients}, ISSN={["2165-0497"]}, DOI={10.1128/spectrum.01031-21}, abstractNote={While the abundance of specific ɣ-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. ABSTRACT This study investigated the ability of ɣ-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 ɣ-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 ɣ-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 ɣ-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 ɣ-proteobacteria were inhibited in CJM supplemented with a low salt fermentation cover brine containing calcium chloride, acetic acid and potassium sorbate. The ɣ-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 ɣ-proteobacteria in CJM is not replicated in cucumber fermentation. While the ɣ-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 ɣ-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 ɣ-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 ɣ-proteobacteria to proliferate and reduce pH to 4.47 ± 0.12. However, ɣ-proteobacteria are unable to proliferate in cucumber fermentation. Control of ɣ-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.}, journal={MICROBIOLOGY SPECTRUM}, author={Rothwell, Madison A. R. and Zhai, Yawen and Pagan-Medina, Christian G. and Perez-Diaz, Ilenys M.}, year={2022}, month={May} }
 @article{zhai_perez-diaz_2021, title={Identification of potential causative agents of the CO2-mediated bloater defect in low salt cucumber fermentation}, volume={344}, ISSN={["1879-3460"]}, DOI={10.1016/j.ijfoodmicro.2021.109115}, abstractNote={Development of bloater defect in cucumber fermentations is the result of carbon dioxide (CO2) production by the indigenous microbiota. The amounts of CO2 needed to cause bloater defect in cucumber fermentations brined with low salt and potential microbial contributors of the gas were identified. The carbonation of acidified cucumbers showed that 28.68 ± 6.04 mM (12%) or higher dissolved CO2 induces bloater defect. The microbiome and biochemistry of cucumber fermentations (n = 9) brined with 25 mM calcium chloride (CaCl2) and 345 mM sodium chloride (NaCl) or 1.06 M NaCl were monitored on day 0, 2, 3, 5, 8, 15 and 21 using culture dependent and independent microbiological techniques and High-Performance Liquid Chromatography. Changes in pH, CO2 concentrations and the incidence of bloater defect were also followed. The enumeration of Enterobacteriaceae on Violet Red Bile Glucose agar plates detected a cell density of 5.2 ± 0.7 log CFU/g on day 2, which declined to undetectable levels by day 8. A metagenomic analysis identified Leuconostocaceae in all fermentations at 10 to 62%. The presence of both bacterial families in fermentations brined with CaCl2 and NaCl coincided with a bloater index of 24.0 ± 10.3 to 58.8 ± 23.9. The prevalence of Lactobacillaceae in a cucumber fermentation brined with NaCl with a bloater index of 41.7 on day 5 suggests a contribution to bloater defect. This study identifies the utilization of sugars and malic acid by the cucumber indigenous Lactobacillaceae, Leuconostocaceae and Enterobacteriaceae as potential contributors to CO2 production during cucumber fermentation and the consequent bloater defect.}, journal={INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY}, author={Zhai, Yawen and Perez-Diaz, Ilenys M.}, year={2021}, month={Apr} }
 @article{zhai_perez-diaz_2020, title={Contribution of Leuconostocaceae to CO 2-mediated bloater defect in cucumber fermentation *}, volume={91}, ISSN={["1095-9998"]}, DOI={10.1016/j.fm.2020.103536}, abstractNote={Fermented cucumber bloater defect, caused by the accumulation of microbiologically produced carbon dioxide (CO2), creates significant economic losses for the pickling industry. The ability of Leuconostocaceae, indigenous to cucumber, to grow and produce CO2 during a fermentation and cause bloater defect was evaluated. Leuconostocaceae grew and produced over 40% CO2 in cucumber juice medium, used as a model for cucumber fermentation. The inoculation of Leuconostocaceae to 5 Log CFU/g in cucumber fermentations brined with 25 mM calcium chloride and 6 mM potassium sorbate resulted in no significant differences in bloater defect, colony counts from MRS and VRBG agar plates or the fermentation biochemistry; suggesting an inability of the inoculated bacterial species to prevail in the bioconversion. Acidified cucumbers were subjected to a fermentation inoculated with a Leuconostoc lactis starter culture after raising the pH to 5.9 ± 0.4. CO2 was produced in the acidified cucumber fermentations to 13.6 ± 3.5% yielding a bloater index of 21.3 ± 6.4; while 8.6 ± 0.8% CO2 and a bloater index of 5.2 ± 5.9 were observed in the non-inoculated control jars. Together the data collected demonstrate that Leuconostocaceae can produce enough CO2 to contribute to bloater defect, if not outcompeted by the leading lactic acid bacteria in a cucumber fermentation.}, journal={FOOD MICROBIOLOGY}, author={Zhai, Yawen and Perez-Diaz, Ilenys M.}, year={2020}, month={Oct} }
 @misc{zhai_perez-diaz_diaz_2018, title={Viability of commercial cucumber fermentation without nitrogen or air purging}, volume={81}, ISSN={["0924-2244"]}, DOI={10.1016/j.tifs.2018.05.017}, abstractNote={Bloated cucumber defect, resulting from the accumulation of the biologically produced carbon dioxide (CO2) in the fruit, reduces yield and economic gains for the pickling industry worldwide. It was the aim of this review to identify commonalities among effective strategies to reduce bloater defect and determine the theoretical viability of commercial cucumber fermentations without bloater defect and/or purging. This article summarizes the known causes of fermented cucumber bloating defect, including sources of CO2, and the strategies developed to mitigate the production of the carbonic gas such as controlled fermentation, inoculation of selected starter cultures, cover brine acidification and reformulation and the application of air or nitrogen purging. It was understood that microbial activity during fermentation, cucumber tissue respiration, as well as the pressure in the fruits and fermentation tanks, ambient temperature and cover brine composition, impact the levels of dissolved CO2 in the system. Although the biological conversion of oxygen to CO2 reduces the cucumbers internal gas pressure, the dissipation of the gas from the tissue is reduced by brining. Once the gas accumulates in the cucumber tissue in concentrations high enough to displace it, the irreversible formation of hollow cavities or bloaters occurs. Residual CO2 is produced by acid-preserved cucumbers, presumably by tissue respiration, which results in the absence of bloating. Thus, microbial activity seems to contribute most of the CO2 needed for cucumbers to bloat. It is speculated that colonization of the internal cucumber tissue by indigenous microbes, in particular aerobic gram-negative bacteria, results in the localized production of CO2 causing bloating defect early in the fermentation. It is concluded that effective manipulation of the microbiota, reduction of dissolved oxygen levels and the use of adequately selected starter cultures may enable cucumber fermentations of acceptable quality without purging.}, journal={TRENDS IN FOOD SCIENCE & TECHNOLOGY}, author={Zhai, Yawen and Perez-Diaz, Ilenys M. and Diaz, Joscelin T.}, year={2018}, month={Nov}, pages={185–192} }
 @article{zhai_pérez-díaz_diaz_lombardi_connelly_2017, title={Evaluation of the use of malic acid decarboxylase-deficient starter culture in NaCl-free cucumber fermentations to reduce bloater incidence}, volume={124}, ISSN={1364-5072}, url={http://dx.doi.org/10.1111/jam.13625}, DOI={10.1111/jam.13625}, abstractNote={Accumulation of carbon dioxide (CO2) in cucumber fermentations is known to cause hollow cavities inside whole fruits or bloaters, conducive to economic losses for the pickling industry. This study focused on evaluating the use of a malic acid decarboxylase (MDC)‐deficient starter culture to minimize CO2 production and the resulting bloater index in sodium chloride‐free cucumber fermentations brined with CaCl2.}, number={1}, journal={Journal of Applied Microbiology}, publisher={Wiley}, author={Zhai, Y. and Pérez-Díaz, I.M. and Diaz, J.T. and Lombardi, R.L. and Connelly, L.E.}, year={2017}, month={Dec}, pages={197–208} }