Clostridium botulinum Toxin Formation in Romaine Lettuce and Shredded Cabbage: Effect of Storage and Packaging Conditions

1995 ◽  
Vol 58 (6) ◽  
pp. 624-627 ◽  
Author(s):  
RUTH L. PETRAN ◽  
WILLIAM H. SPERBER ◽  
ARTHUR B. DAVIS
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Toxin Production ◽  
Romaine Lettuce ◽  
Toxin Detection ◽  
Storage Method ◽  
Fresh Cut ◽  
Storage Temperatures ◽  
Toxin Formation

The potential for growth and toxin production by Clostridium botulinum spores was investigated in samples of fresh-cut romaine lettuce and shredded cabbage packaged in vented and nonvented flexible pouches at storage temperatures of 4.4, 12.7, and 21°C for up to 28 days. One hundred grams of cut, washed romaine lettuce and cabbage were placed in polyester bags. Approximately 104 heat-shocked spores were added per package. Before sealing, half the packages were vented. No toxin was detected with either storage method at 4.4 and 12.7°C. In nonvented pouches of romaine lettuce at 21°C, spores grew and produced toxin after 14 days. Romaine samples at 21°C in vented pouches became toxic after 21 days, indicating formation of anaerobic microenvironments within the pouches. In nonvented packages of cabbage at 21°C, toxin was detected after 7 days. All toxin-positive samples were judged to be inedible prior to toxin detection.

Effect of Equilibrated pH and Indigenous Spoilage Microorganisms on the Inhibition of Proteolytic Clostridium botulinum Toxin Production in Experimental Meals under Temperature Abuse

2017 ◽  
Vol 80 (8) ◽  
pp. 1252-1258 ◽  
Author(s):  
Max C. Golden ◽  
Brandon J. Wanless ◽  
Jairus R. D. David ◽  
D. Scott Lineback ◽  
Ryan J. Talley ◽  
...  
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Storage Temperature ◽  
Phase 1 ◽  
Toxin Production ◽  
Brussels Sprouts ◽  
The Impact ◽  
And Storage ◽  
Spoilage Microorganisms ◽  
Toxin Formation

ABSTRACT Clostridium botulinum is a foreseeable biological hazard in prepared refrigerated meals that needs to be addressed in food safety plans. The objective of this study was to evaluate the effect of product composition and storage temperature on the inhibition of botulinum toxin formation in nine experimental meals (meat, vegetable, or carbohydrate based). Treatments were inoculated with proteolytic C. botulinum, vacuum packaged, cooked at 90°C for 10 min, and assayed for botulinum toxin in samples stored at 25°C for up to 96 h for phase 1, or at 25°C for 12 h and then transferred to 12.5°C for up to 12 and 6 weeks in phases 1 and 2, respectively. For phase 1, none of the treatments (equilibrated pH 5.8) supported toxin production when stored at 25°C for 48 h, but toxin production was observed in all treatments at 72 h. For the remaining experiments with storage at 12.5°C, toxin production was dependent on equilibrated pH, storage time, and growth of indigenous spoilage microorganisms. In phase 1, no gross spoilage and no botulinum toxin was detected for any treatment (pH ≤5.8) stored at 12.5°C for 12 weeks. In phase 2, gross spoilage varied by commodity, with the brussels sprouts meal with pH 6.5 showing the most rapid spoilage within 2 weeks and botulinum toxin detected at 5 and 6 weeks for the control and cultured celery juice treatments, respectively. In contrast, spoilage microbes decreased the pH of a pH 5.9 beef treatment by 1.0 unit, potentially inhibiting C. botulinum through 6 weeks at 12.5°C. None of the other treatments with pH 5.8 or below supported toxin production or spoilage. This study provides validation for preventive controls in refrigerated meals. These include equilibrated product pH and storage temperature and time to inhibit toxin formation by proteolytic C. botulinum, but the impact of indigenous microflora on safety and interpretation of challenge studies is also highlighted.

Factors Inhibiting the Growth and Toxin Formation of Clostridium botulinum Types A and B in “Tsuyu” (Japanese noodle soup)

1990 ◽  
Vol 53 (12) ◽  
pp. 1025-1032 ◽  
Author(s):  
HIROYUKI IMAI ◽  
KATSUNORI OSHITA ◽  
HIKOTAKA HASHIMOTO ◽  
DANJI FUKUSHIMA
Keyword(s):  
Botulinum Toxin ◽  
Salt Concentration ◽  
Clostridium Botulinum ◽  
Incubation Temperature ◽  
Toxin Production ◽  
Distilled Water ◽  
Naoh Solution ◽  
Toxin Formation ◽  
Positive Controls

The objective of this study was to evaluate the safety of five different “tsuyu” (Japanese noodle soups) and the effects of incubation temperature, salt concentration, and pH by pursuing the fate of Clostridium botulinum types A and B spores inoculated into them. C. botulinum did not produce toxin in three concentrated types (pH 5.0–5.3, NaCl 8–12%) of “tsuyu” within 3 months at 30°C and in two ready-to-use types (pH 4.83–4.92, NaCl 3.95–4.05) within 6 months at 20 and 30°C. In addition to the “tsuyu” products, positive controls of ready-to-use “tsuyu” with pH-salt combinations adjusted with a sterile NaOH solution to pH 7.0, 6.5, 6.0, and 5.5 and diluted with sterile distilled water to 3, 2, and 1% salt concentrations were included in this study. Growth and toxin production by C. botulinum in positive controls were dependent upon incubation temperature, pH, and salt concentration. That is, no botulinum toxin would occur in “tsuyu” with 4% salt at pH below 6.5 at 20°C; with 1% salt at pH below 5.0 at 30°C; 2 or 3% salt at pH below 5.5 at 30°C; 4% salt at pH below 6.0 incubated at 30°C, even if contaminated with the spores.

Effect of Cultured Celery Juice, Temperature, and Product Composition on the Inhibition of Proteolytic Clostridium botulinum Toxin Production

2017 ◽  
Vol 80 (8) ◽  
pp. 1259-1265 ◽  
Author(s):  
Max C. Golden ◽  
Brandon J. Wanless ◽  
Jairus R. D. David ◽  
Bala Kottapalli ◽  
D. Scott Lineback ◽  
...  
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Storage Time ◽  
Toxin Production ◽  
Cold Chain ◽  
Effect Of Temperature ◽  
Product Composition ◽  
Challenge Study ◽  
Fail Safe ◽  
Toxin Formation

ABSTRACTClostridium botulinum may be of concern in prepared refrigerated meals, for which strict cold chain management cannot be guaranteed. This study evaluated the effect of temperature, product composition, and cultured celery juice powder (CCJP) as a source of nitrite on the inhibition of botulinum toxin formation in two experimental (meat- and vegetable-based) prepared meals. Data obtained from the challenge study were compared with a published mathematical model to determine whether the model is fail-safe with regard to the tested meals. Treatments were inoculated with proteolytic C. botulinum, vacuum packaged, cooked at 90°C for 10 min, and assayed for botulinum toxin at appropriate intervals in samples stored at 10, 15, or 20°C for up to 8 weeks. None of the treatments stored at 10°C for 8 weeks supported toxin production by proteolytic C. botulinum. The addition of CCJP delayed toxin production by 1 and 3 weeks in cauliflower potatoes and in Dijon pork, respectively, stored at 15°C. Toxin production was delayed by 1 week at 20°C when CCJP was added to the cauliflower potatoes. This study found that the predictive model was fail-safe but was overly conservative for the experimental meals described. Finally, this study confirms that product composition, the addition of nitrite via CCJP, storage time, and temperature play important roles in the inhibition of toxin formation by proteolytic C. botulinum.

Clostridium botulinum Toxin Production in Relation to Spoilage of Atlantic Salmon (Salmo salar) Packaged in Films of Varying Oxygen Permeabilities and with Different Atmospheres

2015 ◽  
Vol 78 (11) ◽  
pp. 2006-2018 ◽  
Author(s):  
MARILYN C. ERICKSON ◽  
LI M. MA ◽  
MICHAEL P. DOYLE
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Transmission Rate ◽  
Toxin Production ◽  
The United States ◽  
Anaerobic Growth ◽  
Commercial Application ◽  
Packaging Films ◽  
Oxygen Transmission Rate ◽  
Storage Temperatures

Shelf life of fish packaged under modified atmosphere (MA) is extended, but within the United States, commercial application of MA with impermeable packaging films is restricted due to concerns that botulinum toxin production would precede spoilage when contaminated fish are held at abusive storage temperatures. Use of semipermeable packaging films has been advocated; however, previous studies are inconclusive in determining the oxygen transmission rate (OTR) of a film that is needed to achieve an acceptable margin of safety (i.e., toxin production occurs only after spoilage). This study was conducted to determine the influence of OTR (target OTRs of 3 to 15,000) on the development of spoilage volatiles and toxin in salmon inoculated with type E Clostridium botulinum and subjected to air, vacuum, or 75:25 CO2:N2 MA and storage temperatures of 4, 8, 12, or 16°C. The most dominant headspace volatile peak that was produced during spoilage of samples at 4, 8 or 12°C was a peak, having a Kovats retention index (KI) of 753, and at which external standards of 2- or 3-methyl 1-butanol also eluted. Under anaerobic conditions, both the aerobic microbial populations and the size of the KI 753 spoilage peak were less in inoculated samples compared with uninoculated samples. C. botulinum–inoculated samples that were stored at 12 or 16°C under conditions favorable for anaerobic growth were also characterized by a KI 688 peak. Using a previously developed model that related the percentage of elderly consumers who would prepare a sample having the KI 753 spoilage peak of a specific size, it was determined that for salmon packaged with 3 or 3,000 OTR films under any atmosphere and stored at 12 or 16°C, 2 to 61% of the consumers could potentially prepare toxin-contaminated samples. Hence, when abusive storage conditions are suspected, the fish should not be consumed.

scholarly journals Effects of Modified Atmosphere Packaging on Toxin Production by Clostridium botulinum in Raw Aquacultured Summer Flounder Fillets (Paralichthys dentatus)

2007 ◽  
Vol 70 (5) ◽  
pp. 1159-1164 ◽  
Author(s):  
FLETCHER M. ARRITT ◽  
JOSEPH D. EIFERT ◽  
MICHAEL L. JAHNCKE ◽  
MERLE D. PIERSON ◽  
ROBERT C. WILLIAMS
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Modified Atmosphere Packaging ◽  
Toxin Production ◽  
Plate Count ◽  
Modified Atmosphere ◽  
Paralichthys Dentatus ◽  
Microbial Spoilage ◽  
Fish Fillets ◽  
Toxin Formation

Packaging fishery products under vacuum atmosphere packaging (VAC) and modified atmosphere packaging (MAP) conditions can significantly extend the shelf life of raw, refrigerated fish products. There is considerable commercial interest in marketing VAC and MAP refrigerated (never frozen) raw fish fillets. The objective of this study was to determine if Clostridium botulinum toxin development precedes microbiological spoilage in raw, refrigerated flounder fillets. Aquacultured flounder (Paralichthys dentatus) individual fish fillets either were packed with a film having an oxygen transmission rate (OTR) of 3,000 cm3 m−2 24 h−1 at 22.8°C or were vacuum packaged or packaged under 100% CO2 with a film having an OTR of 7.8 cm3 m−2 24 h−1 at 21.1°C and were stored at 4 and 10°C. Samples were analyzed by aerobic plate count (APC) for spoilage and qualitatively for botulinum toxin with a mouse bioassay. The results demonstrate that flounder fillets (4°C) packaged with a film having an OTR of 3,000 were microbiologically spoiled (APC, >107 CFU/g) on day 15, but there was no toxin formation, even after 35 days of storage. However, at 10°C, toxin production occurred (day 8), but it was after microbial spoilage and absolute sensory rejection (day 5). Vacuum-packaged fillets and 100% CO2 fillets (4°C) packaged with a film having an OTR of 7.8 were toxic on days 20 and 25, respectively, with microbial spoilage (APC, >107 CFU/g) not occurring during the tested storage period (i.e., >35 days). At 10°C, in vacuum-packaged flounder, toxin formation coincided with microbiological spoilage (days 8 to 9). In the 100% CO2-packaged fillets, toxin formation occurred on day 9, with microbial spoilage occurring on day 15. This study indicates that films with an OTR of 3,000 can be used for refrigerated fish fillets and still maintain the safety of the product.

Growth and Toxin Production by Clostridium botulinum on Inoculated Fresh-Cut Packaged Vegetables

1998 ◽  
Vol 61 (3) ◽  
pp. 324-328 ◽  
Author(s):  
J. W. AUSTIN ◽  
K. L. DODDS ◽  
B. BLANCHFIELD ◽  
J. M. FARBER
Keyword(s):  
Botulinum Toxin ◽  
Spore Germination ◽  
Low Temperatures ◽  
Clostridium Botulinum ◽  
Toxin Production ◽  
Modified Atmosphere ◽  
Potential Growth ◽  
Different Temperatures ◽  
Fresh Cut ◽  
Germination And Growth

To determine the safety of fresh-cut vegetables packaged in modified atmosphere, challenge studies using both nonproteolytic and proteolytic strains of Clostridium botulinum were performed with a variety of fresh-cut packaged salads and vegetables stored at different temperatures. When vegetables were inoculated with spores of C. botulinum and incubated in low-O2 atmospheres, spore germination and growth and toxin production were observed. Botulinum toxin was produced by proteolytic types A and B on onion, butternut squash, rutabaga, salad, and stir-fry vegetables. Nonproteolytic C. botulinum produced toxin on butternut squash and salad. Nonproteolytic C. botulinum was capable of producing neurotoxin at temperatures as low as 5°C, whereas proteolytic strains produced neurotoxin at 15°C and higher. Although most samples were visibly spoiled before detection of botulinum toxin, samples of butternut squash and onion remained acceptable after detection of toxin. The strict maintenance of low temperatures (<5°C) is recommended in order to control the potential growth of C. botulinum on fresh-cut vegetables packaged in a modified atmosphere.

Effect of Headspace Oxygen Concentration on Growth and Toxin Production by Proteolytic Strains of Clostridium botulinum

1992 ◽  
Vol 55 (1) ◽  
pp. 23-27 ◽  
Author(s):  
R. C. WHITING ◽  
K. A. NAFTULIN
Keyword(s):  
Clostridium Botulinum ◽  
Critical Level ◽  
Toxin Production ◽  
Inoculum Size ◽  
Oxygen Levels ◽  
Ph Values ◽  
Headspace Oxygen ◽  
Series Of Experiments ◽  
Germination And Growth ◽  
Toxin Formation

A series of experiments was conducted to determine growth and toxin formation by proteolytic strains of Clostridium botulinum in broth media that have known pH values (5–7), NaCl concentrations (0–4%), and controlled oxygen-nitrogen atmospheres. Lower pH and higher NaCl levels inhibited growth and toxin production by vegetative cells, but 15% oxygen in the headspace was insufficient for inhibition in all media. When spores were used as inocula and the tubes were gas flushed, outgrowth and toxin production generally occurred only under a 1% or less oxygen atmosphere. Occurrence of growth and toxin was favored by high pH and low NaCl levels and was related to spore inoculum size. Spores were also inoculated into a mixed fermenter with controlled oxygen levels in the headspace. Times to measurable turbidity increased with greater oxygen levels from 36 h at 0.005% O2 to 109 h at 0.7% O2; however, growth rates were unaffected by headspace oxygen levels. No toxin was observed with 0.9% O2, further demonstrating that the critical level of oxygen for germination and growth is approximately 1%.

Conservative Prediction of Time to Clostridium botulinum Toxin Formation for Use with Time-Temperature Indicators To Ensure the Safety of Foods

1998 ◽  
Vol 61 (9) ◽  
pp. 1154-1160 ◽  
Author(s):  
GUY E. SKINNER ◽  
JOHN W. LARKIN
Keyword(s):  
Botulinum Toxin ◽  
Empirical Relationship ◽  
Critical Time ◽  
Food Product ◽  
Toxin Production ◽  
Storage Conditions ◽  
Potential Health ◽  
And Storage ◽  
Infinite Set ◽  
Toxin Formation

Integrating-type time-temperature indicators (TTIs) may be utilized to warn food processors and consumers about storage conditions that may have rendered a food potentially hazardous. As an example of how integrated TTIs could be manufactured to emulate an infinite set of time-temperature situations, a set of conditions which have supported C. botulinum growth and toxin production was compiled. The time-temperature curve representing conservative times required for toxin formation was constructed with data from literature relating to toxin formation as a function of temperature in any media or food product. This set of critical time-temperature data is fit by a conservative empirical relationship that can be used to predict combinations of incubation times and storage temperatures that represent a potential health risk from C. botulinum in foods. A TTI could be constructed to indicate deviation from such a given set of conditions to bring attention to foods that may have been exposed to potentially hazardous temperatures with respect to C. botulinum toxin formation.

Sign in / Sign up

Export Citation Format

Share Document