Development of a DNA Array for the Simultaneous Detection and Identification of Sugar Thick Juice Bacterial Contaminants

Despite the use of generally accepted good storage practices, sugar thick juice degradation caused by microbiological contamination occasionally occurs, causing considerable financial loss. In this study, a DNA array was developed for simultaneous detection and identification of the most prominent microflora present during thick juice storage, which may cause degradation of the thick juice. Specific oligonucleotides were developed for several bacterial taxa, including the genera Bacillus, Kocuria, Staphylococcus and Tetragenococcus and the species Aerococcus viridans, Leuconostoc mesenteroides and Tetragenococcus halophilus. The DNA array was validated using both pure cultures and industrial samples. In addition, comparisons were made between the developed array, PCR assays specifically targeting the thick juice contaminants and classical microbial platings. The array was found to be reliable and sensitive enough to detect and identify the target bacteria. In addition, the array was used to monitor the target microbial populations in thick juice during long-term storage and degradation. Results are discussed in relation to DNA stability in thick juice.     

Predominance of Tetragenococcus halophilus as the cause of sugar thick juice degradation

The industrial storage of sugar thick juice was simulated on a laboratory scale. Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis and the application of Clone Libraries in parallel with classical microbiology were used to study the bacterial diversity and all revealed a dominance (>99%) of Tetragenococcus halophilus during storage. The degradation of thick juice correlated with the appearance of L-lactic acid and high concentrations of T. halophilus. In addition, pure cultures of T. halophilus induced degradation of sterile thick juice. A specific PCR was developed to detect T. halophilus and industrial thick juice samples from Belgium, Germany and France all contained T. halophilus, suggesting a consistent association of this organism with thick juice. T. halophilus has been known only as a halophile thus far, and this report is the first to show an association of this organism with a sugar-rich environment.     

D-alanine in Fruit Juices: A Molecular Marker of Bacterial Activity, Heat Treatments and Shelf-life

Apple, grapefruit, orange, peach, pear, pineapple and other commercial juice samples were investigated by GC (chiral phase) and HPLC (chiral eluent) to detect D-amino acids as molecular markers of bacterial activity, heat treatments, and shelf life. The development of D-amino acids was followed in grapefruit juice samples inoculated with bacteria (Lactobacillus plantarun), or yeasts (Saccharomyces cereviae). Significant amounts of free D-alanine were found only in juices affected by bacterial contamination. The content of D-alanine was not dependent upon pasteurization or sterilization treatments and did not change during the shelf-life of the products. D-alanine may be considered as a marker of bacterial contamination occurring before or during juice processing.     

Strategies to reduce mycotoxin levels in maize during storage: a review

Maize (Zea mays L.) is one of the main cereals as a source of food, forage and processed products for industry. World production is around 790 million tonnes of maize because as a staple food it provides more than one-third of the calories and proteins in some countries. Stored maize is a man-made ecosystem in which quality and nutritive changes occur because of interactions between physical, chemical and biological factors. Fungal spoilage and mycotoxin contamination are of major concern. Aspergillus and Fusarium species can infect maize pre-harvest, and mycotoxin contamination can increase if storage conditions are poorly managed. Prevention strategies to reduce the impact of mycotoxin in maize food and feed chains are based on using a hazard analysis critical control point systems (HACCP) approach. To reduce or prevent production of mycotoxins, drying should take place soon after harvest and as rapidly as feasible. The critical water content for safe storage corresponds to a water activity (a _w) of about 0.7. Problems in maintaining an adequately low a _w often occur in the tropics where high ambient humidity make the control of commodity moisture difficult. Damage grain is more prone to fungal invasion and, therefore, mycotoxin contamination. It is important to avoid damage before and during drying, and during storage. Drying maize on the cob before shelling is a very good practice. In storage, many insect species attack grain and the moisture that can accumulate from their activities provides ideal conditions for fungal activity. To avoid moisture and fungal contamination, it is essential that the numbers of insects in stored maize should be kept to a minimum. It is possible to control fungal growth in stored commodities by controlled atmospheres, preservatives or natural inhibitors. Studies using antioxidants, essential oils under different conditions of a _w, and temperature and controlled atmospheres have been evaluated as possible strategies for the reduction of fungal growth and mycotoxin (aflatoxins and fumonisins) in stored maize, but the cost of these treatments is likely to remain prohibitive for large-scale use.     

In silico guided pre-treatment of sugarcane juice with natural inhibitors of polyphenol oxidase (PPO) is a new and effective strategy for development of spray-dried formulation

Sugarcane juice is a popular beverage and is also processed to produce sugar. The polyphenol oxidase (PPO) in sugarcane juice causes enzymatic browning and makes the process of sugar production complex and cumbersome. Storage of sugarcane juice is also hampered by the high sugar content and rapid microbial fermentation. The present research assessed the potential of lemon juice (LJ) and ginger extract (GE) as natural inhibitors of PPO. Enzyme kinetics and the mechanism of inhibition of LJ and GE were studied. Primary investigation was carried out using molecular docking approach to assess the inhibitory potential of LJ and GE and to determine the nature of interaction between the enzyme and inhibitors. Extracts were used as inhibitors and studies revealed that both reduced the PPO activity. Subsequently, pure bioactive inhibitors such as ascorbic acid, citric acid, and 6-shogaol present in these natural extracts were used to study the mode of inhibition of PPO. Citric acid decreased PPO activity by lowering pH, while ascorbic acid was found to be a competitive inhibitor of PPO with a K_i of 75.69 µM. The proportion of LJ and GE required in sugarcane juice was optimized on the basis of browning index and sensory acceptance. Further, the sugarcane cane juice after inhibition of PPO under optimized conditions was spray dried and evaluated for reconstitution properties. The product formulated in the present study is a new and effective approach to address quality-compromising issues associated with long-term storage of cane juice.     

Effect of different levels of CO2 on the antioxidant content and the polyphenol oxidase activity of ‘Rocha’ pears during cold storage

Pears (Pyrus communis L. cv. ‘Rocha’) were exposed to air or controlled atmosphere (CA) containing various concentrations of CO₂: 0, 0.5 and 5 kPa, all with 2 kPa O₂. After 4 months of storage at 2 °C, the fruits were transferred to air at room temperature, and assessed in terms of soluble solids, titratable acidity, pH, incidence of brown heart and flesh browning, phenolic content, vitamin C content and polyphenol oxidase activity. By 4 months of storage, soluble solids and pH increased, and acidity decreased relative to harvest, but no differences were detected between pears stored under air or any of the CA tested. Higher contents of hydroxycinnamic derivatives and flavan‐3‐ols in the peel than in the flesh were recorded. However, the content of arbutin was higher in the flesh than in the peel, whereas flavonols were only detected in the peel. In general, hydroxycinnamic derivatives and flavonols were stable throughout storage, but flavan‐3‐ols decreased in concentration under air or CA. Arbutin was the only phenolic compound that increased in concentration as time elapsed. No clear relation was found between the storage conditions tested and the phenolic concentration in pears. Regarding ascorbic acid (AA) and dehydroascorbic acid (DHA), their concentrations were higher in the peel than in the flesh. Furthermore, AA and DHA were strongly affected by storage: the former decreased, whereas the latter increased in content. A decrease in PPO activity was apparent after harvest and during storage, particularly under higher levels of CO₂. The combination 2 kPa O₂ + 5 kPa CO₂ increased the incidence of internal disorders (viz. brown heart and flesh browning) after storage. Copyright © 2005 Society of Chemical Industry     

Influence of combined hot air and UV-C treatment on the antioxidant system of minimally processed broccoli (Brassica oleracea L. var. Italica)

Broccoli develops a fast senescence that deteriorates the nutritional quality of the product. In this work, a combined treatment with hot air and UV-C were applied to minimally processed broccoli florets to investigate their effects on the antioxidant system during storage at 20 °C. Application of UV-C and heat enhanced by approximately 13% the levels of total antioxidants immediately after treatment. These levels were lower than initials in both control and treated samples. Nevertheless, higher values (ranged from 12 to 50%) were also detected in treated samples during storage. In general, higher values of antioxidants were correlated with elevated levels of phenols (from 10 to 17%) and ascorbic acid (from 11 to 17%) in treated florets. The increment in the content of phenolics in treated broccolis was correlated to an increased phenylalanine ammonia lyase (PAL) activity. The combined treatment also affects the activity of enzymes involved in removal of reactive oxygen species. An increase of about 50% in superoxide dismutase activity was detected after treatment. During the first days of storage this higher activity was about 40%. Enzymes that remove H₂O₂ like catalase and ascorbate peroxidase showed an enhanced activity toward the end of storage. The combined treatment diminished the peroxidase (POX) activity approximately 60% after the treatment and near 50% after two days of storage, suggesting a minor role of this enzyme in detoxification of H₂O₂. In conclusion, combined treatment may contribute to enhance the protection against oxidative molecules not only by increasing levels of phenolics and ascorbic acid but also by enhancing the activity of enzymes involved in removing reactive oxygen species.     

Soluble Sugar Content and Metabolism as Related to the Heat-Induced Chilling Tolerance of Loquat Fruit During Cold Storage

The effects of high-temperature, short-time hot air treatment (45 °C for 3 h) on soluble sugar metabolism and chilling tolerance in loquat fruit stored at 5 °C for 5 weeks were investigated. Heat treatment significantly reduced chilling severity, as evidenced by lower firmness and internal browning and higher levels of extractable juice. Meanwhile, this treatment accelerated the activities of acid invertase, neutral invertase, sucrose phosphate synthase and sucrose synthase during storage. However, sucrose degradation was predominant, which caused lower levels of sucrose and higher levels of glucose and fructose in the heat-treated group. In addition, the ascorbate acid content and the activities of ascorbate peroxidase and glutathione reductase in the heat-treated fruit were much higher than those in control fruit, resulting in lower levels of hydrogen peroxide (H₂O₂) and malondialdehyde and decreased membrane permeability as well as a higher unsaturated/saturated fatty acid ratio at the end of storage. Our results suggest that the increased levels of reducing sugars, especially those of glucose, may induce the ascorbate–glutathione cycle activity to scavenge for H₂O₂, whose content relates to the heat-induced chilling tolerance of loquat fruit.     

Effects of modified atmosphere and vacuum packaging on the growth of spoilage and inoculated pathogenic bacteria on fresh poultry

Fresh chicken breast meats inoculated withYersinia enterocolitica andAeromonas hydrophila were packaged in glass jars either containing different compositions of modified atmospheres (MA) (100% CO₂; 80% CO₂/20% N₂), or in vacuo or containing air, and were stored at 3±1°C and 8±1°C. The changes in gas composition as well asY. enterocolitica, A. hydrophila, total aerobic bacterial, total psychrotrophic, Lactobacilli and Enterobacteriaceae counts were determined after 0,1,3,7,9,11 and 14 days of storage. The results show that while the growth ofY. enterocolitica andA. hydrophila were retarded following MA storage, the pathogens were capable of growth in MA and vacuum storage at both temperatures, for the inoculation levels studied. For total aerobic bacterial counts, there were no differences between the values for chicken breast meats kept in different atmospheres. Being packaged in CO₂ had the greatest inhibitory effect on the growth of psychrotrophic aerobic bacteria during the first 3 days. Lactic acid bacteria levels of samples stored in MA conditions and in vacuo increased rapidly when compared to those levels of samples stored in air. It was also found that the effect of MA storage increased at 3±1°C.     

Influence of storage environment on maize grain: CO2 production,dry matter losses and aflatoxins contamination

Poor storage of cereals, such as maize can lead to both nutritional losses and mycotoxin contamination. The aim of this study was to examine the respiration of maize either naturally contaminated or inoculated with Aspergillus flavus to examine whether this might be an early and sensitive indicator of aflatoxin (AF) contamination and relative storability risk. We thus examined the relationship between different interacting storage environmental conditions (0.80–0.99 water activity (a_w) and 15–35°C) in naturally contaminated and irradiated maize grain + A. flavus on relative respiration rates (R), dry matter losses (DMLs) and aflatoxin B1 and B2 (AFB1-B2) contamination. Temporal respiration and total CO₂ production were analysed by GC-TCD, and results used to calculate the DMLs due to colonisation. AFs contamination was quantified at the end of the storage period by HPLC MS/MS. The highest respiration rates occurred at 0.95 a_w and 30–35°C representing between 0.5% and 18% DMLs. Optimum AFs contamination was at the same a_w at 30°C. Highest AFs contamination occurred in maize colonised only by A. flavus. A significant positive correlation between % DMLs and AFB1 contamination was obtained (r = 0.866, p < 0.001) in the irradiated maize treatments inoculated with A. flavus. In naturally contaminated maize + A. flavus inoculum loss of only 0.56% DML resulted in AFB1 contamination levels exceeding the EU legislative limits for food. This suggests that there is a very low threshold tolerance during storage of maize to minimise AFB1 contamination. This data can be used to develop models that can be effectively used in enhancing management for storage of maize to minimise risks of mycotoxin contamination.     

Application of Ozonated Dry Ice (ALIGAL™ Blue Ice) For Packaging and Transport in the Food Industry

Abstract: Dry ice is used by meat and poultry processors for temperature reduction during processing and for temperature maintenance during transportation. ALIGAL? Blue Ice (ABI), which combines the antimicrobial effect of ozone (O₃) along with the high cooling capacity of dry ice, was investigated for its effect on bacterial reduction in air, in liquid, and on food and glass surfaces. Through proprietary means, O₃ was introduced to produce dry ice pellets to a concentration of 20 parts per million (ppm) by total weight. The ABI sublimation rate was similar to that of dry ice pellets under identical conditions, and ABI was able to hold the O₃ concentration throughout the normal shelf life of the product. Challenge studies were performed using different microorganisms, including E. coli, Campylobacter jejuni, Salmonella, and Listeria, that are critical to food safety. ABI showed significant (P < 0.05) microbial reduction during bioaerosol contamination (up to 5‐log reduction of E. coli and Listeria), on chicken breast (approximately 1.3‐log reduction of C. jejuni), on contact surfaces (approximately 3.9 log reduction of C. jejuni), and in liquid (2‐log reduction of C. jejuni). Considering the stability of O₃, ease of use, and antimicrobial efficacy against foodborne pathogens, our results suggest that ABI is a better alternative, especially for meat and poultry processors, as compared to dry ice. Further, ABI can potentially serve as an additional processing hurdle to guard against pathogens during processing, transportation, distribution, and/or storage. Practical Application: A new product known as ALIGAL? Blue Ice (ABI) incorporates ozone in dry ice pellets, combining the high cooling capacity of dry ice pellets and the antimicrobial efficacy of ozone. This study evaluated the antimicrobial efficacy of ABI compared to dry ice under potential commercial processing conditions, specifically examining the impact on airborne, surface, moisture, and food contamination. Treatment with ABI resulted in higher reductions of bacteria with airborne contamination, on chicken breast, on surfaces, and in liquid compared to dry ice. Thus, ABI can serve as an additional processing hurdle to guard against pathogens during food processing and storage.     

Fungi and natural incidence of selected mycotoxins in barley rootlets

The storage of barley rootlets is increasingly employed to provide raw material for pig feeding in Brazil. Barley rootlets represent an important feedstuff for animal production due to their high levels of protein and fiber, and low price. However, poor management of raw materials during storage can result in fungal growth, the loss of nutritive substances and contamination by mycotoxins. The aims of this work were (1) to identify fungi associated with barley rootlets used as pig feedstuff raw material, and (2) to identify and quantify selected mycotoxins naturally produced by isolated mycotoxin-producing species in this substrate over a year. Samples were examined for fungal counts and genera distribution. Fumonisin B₁ and aflatoxin B₁ contamination were determined using high pressure liquid chromatography (HPLC). Barley rootlet samples were of low hygienic quality. Although a broad survey was undertaken, low fungal diversity was found. Fusarium verticillioides was the most prevalent species followed by Aspergillus flavus. Despite Aspergillus clavatus being widely associated with high-moisture sprouted grains including brewers’ grains, and causing toxicity to livestock, it was not detected in this work. Although pre-harvest contamination of the barley crop, as in the maize, could occur, the barley might support F. verticillioides/Fusarium proliferatum growth when grain is remoistened during the germination and malting process and it might even continue during storage on pig farms. All samples were positive for fumonisin B₁ whereas aflatoxin B₁ contamination was not detected. It is important to point out the potential risk of fumonisin contamination in barley rootlets used as animal feed. Fusarium toxins are important not so much for their acute effects as for the chronic syndromes reported worldwide. The obtained results reveal the need for periodic monitoring of raw materials to avoid problems in animal production and hazards to animal and human health.     

Evaluation of ethanol fermentation parameters for bioethanol production from sugar beet pulp and juice

The aim of this study was to investigate the efficient utilization of sugar beet pulp, as well as raw, concentrated raw and thick sugar beet juice, for bioethanol production. Different fermentation conditions were examined. The influence of raw material pre‐treatment (pasteurization or sterilization), type of batch culture process (stationary or shaken) as well as the type of Saccharomyces cerevisiae yeast preparation on the yields of the process were studied. Moreover, the fermentation process effectiveness was examined in connection with the quality of the obtained distillates. Sterilization, stationary batch culture and Safdistil C‐70 yeast preparation were identified as the most profitable factors for sugar beet pulp fermentation, providing a high fermentation efficiency and ethanol yield (87.7% of theoretical ethanol yield). Concentrated raw beet juice resulted in a value of 94.2% of theoretical yield, and thick juice a 92.6% yield. The results suggest that bioethanol production from sugar beet pulp and sugar beet juice has promise as an alternative fuel. The raw spirits obtained from the sugar beet juice were characterized as having the lowest quantity of volatile by‐products. Copyright © 2014 The Institute of Brewing & Distilling     

Catalase, Lipoxygenase, and Peroxidase Activities in Cucumber Pickles as Affected by Fermentation, Processing, and Calcium Chloride

Catalase, lipoxygenase, and peroxidase activities were determined in tissues and brines of pickling cucumbers during fermentation, storage, and after processing. Lipoxygenase and catalase were inactivated within three days of exposure to fermentation brines. Peroxidase activity was relatively unaffected by the first three days of brining, but it declined during fermentation, storage, and after processing. CaCl₂ reduced the loss of peroxidase activity in tissue and brine during fermentation and storage. Alum in post-desalting brine reduced peroxidase activity in processed pickles although CaCl₂ ameliorated its efficacy. No activity of catalase, lipoxygenase or peroxidase was detected in cucumber juice extracts fermented by four different lactic acid bacteria.     

Kinetic Modeling of Anthocyanin Degradation and Microorganism Growth during Postharvest Storage of Açai Fruits (Euterpe oleracea)

Abstract: The unavoidable damage of açai (Euterpe oleracea) fruits (AF) during picking leads to microbial contamination and anthocyanin degradation, which prejudice the consumed fruit drink. Thirteen lots of AF (24 kg) from different municipal districts of the Pará State (Brazil) were monitored during a 75‐h‐long storage in the dark at 30 °C for microbial growth, and 7 lots for anthocyanin degradation. On arrival at the laboratory, anthocyanins presented a mean concentration of 828 mg kg⁻¹ fruits with a standard deviation of 323 mg kg⁻¹ fruits whereas mean microbial contamination was 2.64 10⁶ CFU g⁻¹ of dry matter for total mesophilic bacteria, 1.98 10³ MPN g⁻¹ DM for fecal coliforms, and 1.11 10⁵ CFU g⁻¹ DM for moulds and yeasts. Kinetic growth of the microbes could be fitted to a quadratic equation with an unusual rapid growth during the 1st 24 h. The kinetics of anthocyanin degradation fitted a 1st‐order equation. The mean velocity constant of the reaction (k₁) was of 0.0137 h⁻¹ and the mean half‐life (t_½) of the anthocyanins was 50 h. These results indicate that the AF simultaneously suffer extensive anthocyanin degradation and explosive microbial growth during the postharvest period needing a special care. Practical Application: The data presented in this article help the industrial sector of açai (Euterpe oleracea) juice to understand: (1) the rapid kinetics of anthocyanin oxidation and (2) the uncommon kinetics of microorganism growth during a postharvest storage of açai fruits for up to 75 h. Our results allow to predict the global deterioration of açai fruits and pointed out the necessity of blanching the fruits or pasteurizing the juice.     

Chemometrics coupled with ultraviolet spectroscopy: a tool for the analysis of variety,adulteration, quality and ageing of apple juices

This study demonstrates the use of UV spectroscopy (UV) in combination with chemometrics as a simple and feasible approach for analysis of variety, adulteration, quality and ageing of apple juice. The results show that PCA‐UV is adequate to differentiate apple juice varieties and adulteration. The percentage of the adulterant can be detected by PLSR‐UV with RMSE < 0.7783% and R² > 0.9980. For the evaluation of juice quality, PLSR‐UV (RMSE = 0.2555–2.3448; R² = 0.7276–0.9816) is recommended for the prediction of soluble solids, ascorbic acid, total flavonoids, total sugar and reducing sugar, whilst PCR‐UV (RMSE = 0.0000–2.7426; R² = 0.7073–1.0000) is adequate for the prediction of pH and antioxidant activity. In addition, PLSR‐UV may be used to predict the storage time with RMSE = 0.4681 day and R² = 0.9832. Therefore, UV coupled with chemometrics has potential to be developed as a portable tool for the detection of variety, adulteration, quality and ageing of not only apple juices, but also other fruit and vegetable juices.     

Coupling osmotic dehydration with heat treatment for green papaya impregnated with blackberry juice solution

Slices (1.5 mm thick) of green papaya were impregnated through osmotic dehydration with a blackberry juice–sucrose solution to produce an intermediate moisture product. The effect of processing temperature (T) and sucrose-added molality (m_sucrose) on mass transfer during the operation was assessed, using a response surface methodology (RSM). The RSM was used to model water loss, sugar and anthocyanin gain during the process. Increasing sucrose molality resulted in increasing water loss and sugar gain, but decreasing anthocyanin gain. Water transfer therefore limits anthocyanin impregnation, but not sucrose incorporation. Afterwards, the impact of heat treatment at high temperatures was analysed, using numerical simulation. The conditions of the combined process, designed to achieve an anthocyanin-rich final product, are low sucrose-added molalities (sucrose molality < 1 mol kg^–1) and high processing temperatures (T > 50 ºC) for osmotic dehydration, coupled with high-temperature, short-time (HTST) heat treatments for product stabilisation.     

Laboratory evaluation of previously fermented juice as a fermentation stimulant for lucerne silage

The potential use of forage juice anaerobically stored for 2 days (previously fermented juice, PFJ) as a silage additive was investigated. Lucerne (450 g) was ensiled in laboratory‐scale silos (900 ml capacity) with or without sucrose (20 g kg⁻¹) or PFJ (2.5 ml kg⁻¹). Silages treated with both additives were also made and fermentation quality, in vitro dry matter digestibility and in vitro ruminal NH₃ production were determined. The control silage showed very poor quality with high butyrate and NH₃‐N concentrations. Addition of sucrose significantly increased lactate but failed to inhibit the deterioration by clostridial fermentation. Use of PFJ further increased lactate and greatly reduced butyrate and NH₃‐N production during storage. The combination of sucrose and PFJ was more effective in improving fermentation than either additive on its own. In vitro digestibility appeared to be higher when lucerne was well preserved. Ruminal NH₃ production in vitro was decreased in silages with sucrose and PFJ added at ensiling. This result indicates that use of PFJ can be a promising way to stimulate lactate production in some difficult direct cut silage. The potential of improving fermentation may be comparable to conventional sugar additives. © 1999 Society of Chemical Industry     

Malic Acid as a Source of Carbon Dioxide in Cucumber Juice Fermentations

The degradation of malic acid to lactic acid and CO₂ during fermentation of cucumber juice was investigated. This malolactic reaction was the major source of CO₂ when cucumber juice was fermented by Lactobacillus plantarum. It may also be an important CO₂ source in controlled cucumber fermentations. In addition to CO₂ production, the degradation of malate served to buffer the fermentation and increase sugar utilization. The pH after 7 days’fermentation was 2.8 when 13 mM malic acid was present in the juice and 4.1 with 135 mM malic acid. In the same fermentations, 52% of the sugars were degraded with the low malic acid concentration while complete sugar utilization was observed with the highest malic acid level.     

Antibacterial Activity of Heated Cabbage Juice, S-Methyl-L-Cysteine Sulfoxide and Methyl Methanethiosulfonate

Autoclaved cabbage juice was inhibitory to growth of Staphylococcus aureus. S-Methyl-L-cysteine sulfoxide (SMCSO), autoclaved either together with or separately from nutrient broth, also inhibited the growth of S. aureus, but inhibition was greater when SMCSO was autoclaved separately. Methyl methanethiosulfonate (MMTSO₂), a thermal breakdown product of SMCSO, completely inhibited growth of S. aureus at 10 ppm. MMTSO₂ was formed in both autoclaved samples of cabbage juice and aqueous solution of SMCSO. Thus, evidence indicates that the bacterial inhibitory activity in autoclaved cabbage juice was due to heatinduced formation of MMTSO₂ from SMCSO.