Inhibition of Clostridium botulinum Types A and E Toxin Formation by Sodium Nitrite and Sodium Chloride in Hot-Process (Smoked) Salmon

1982 ◽  
Vol 45 (9) ◽  
pp. 833-841 ◽  
Author(s):  
G. A. PELROY ◽  
M. W. EKLUND ◽  
R. N. PARANJPYE ◽  
E. M. SUZUKI ◽  
M. E. PETERSON

Sodium nitrite and NaCl were evaluated as inhibitors of outgrowth and toxin production by Clostridium botulinum types A and E in abuse-stored (25°C) hot-process salmon. Salmon steaks were brined in NaCl or NaCl plus NaNO2 and inoculated intramuscularly with spores. Steaks were then heated in a simulated hot-smoke process to internal temperatures of 62.8 to 76.7°C (145 to 170°F) for the final 30 min of a 3- to 4-h process, packaged in oxygen-impermeable film and stored at 25°C. During 7 days of storage, toxin production in steaks inoculated with 102 spores per g was inhibited by more than 3.8% water-phase NaCl for type E and 6.1% for type A. Presence of nitrite substantially reduced the salt level required to prevent toxin production. When steaks had more than 100 ppm NaNO2, only 2.5% NaCl inhibited type E toxin production; 150 ppm NaNO2 and 3.5% NaCl inhibited production of type A toxin. When storage time was lengthened to 14 days or the spore inoculum increased to 104 spores per g, more salt and nitrite were required for inhibition. Residual nitrite in samples stored under refrigeration (3.3°C) did not change during 22 days of storage. Under abuse temperature (25°C), residual nitrite decreased to less than 6 ppm by the 14th day in all samples tested regardless of the original nitrite concentration.

2017 ◽  
Vol 80 (8) ◽  
pp. 1259-1265 ◽  
Author(s):  
Max C. Golden ◽  
Brandon J. Wanless ◽  
Jairus R. D. David ◽  
Bala Kottapalli ◽  
D. Scott Lineback ◽  
...  

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.


1987 ◽  
Vol 50 (3) ◽  
pp. 212-217 ◽  
Author(s):  
S. L. CUPPETT ◽  
J. I. GRAY ◽  
J. J. PESTKA ◽  
A. M. BOOREN ◽  
J. F. PRICE ◽  
...  

The effect of salt level and nitrite on botulinal safety of smoked whitefish was investigated. An average water-phase (wp) salt concentration of 4.4% inhibited outgrowth of Clostridium botulinum type E spores (103 spores/g) for over 35 d in temperature-abused (27°C) smoked whitefish. Incorporation of nitrite (220 mg/kg) during brining to the smoked salted (4.4%, wp) whitefish inhibited toxin production for 56 d at 27°C. An average salt concentration of 6.2% (wp), with or without nitrite, totally inhibited toxin production for the duration of the study (83 d). The effect of pH and water activity in temperature-abused smoked whitefish as a means of controlling toxin production by C. botulinum type E spores was evaluated.


1974 ◽  
Vol 27 (4) ◽  
pp. 733-737 ◽  
Author(s):  
L. N. Christiansen ◽  
R. B. Tompkin ◽  
A. B. Shaparis ◽  
T. V. Kueper ◽  
R. W. Johnston ◽  
...  

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 ◽  
...  

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.


1995 ◽  
Vol 58 (6) ◽  
pp. 624-627 ◽  
Author(s):  
RUTH L. PETRAN ◽  
WILLIAM H. SPERBER ◽  
ARTHUR B. DAVIS

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.


1992 ◽  
Vol 55 (1) ◽  
pp. 23-27 ◽  
Author(s):  
R. C. WHITING ◽  
K. A. NAFTULIN

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%.


1987 ◽  
Vol 50 (4) ◽  
pp. 330-336 ◽  
Author(s):  
GENERO W. GARCIA ◽  
CONSTANTIN GENIGEORGIS ◽  
SEPPO LINDROTH

In factorial design experiments we inoculated fresh salmon fillets with a spore pool of 13 nonproteolytic strains of Clostridium botulinum type B, E, and F at 6 levels (10−1 to 104/50 g of fillet), and incubated at 1, 4, 8, 12 and 30°C under modified atmospheres (MA) of vacuum, 100% CO2 and 70% CO2 + 30% air for up to 60 d. The earliest time we detected toxin in the fillets at 30, 12 and 8°C, irrespective of MA, was after 1, 3–9 and 6–12 d of storage and required 100–103, 101–103, 101–102 spores/fillet. The probability (P) of toxin production was significantly (P<0.05) affected by temperature (T), MA storage time (ST), MA × T, MA × ST and T × ST. Only type B toxin was detected in the toxic fillets. No toxin was detected in fillets stored at 4°C for up to 60 d. Toxin detection coincided with spoilage at 30°C, but preceded spoilage at 8 and 12°C, and followed spoilage at 4°C. Using linear and logistic regression analysis, best fit equations were derived relating the length of the lag phase and P of toxin production to T, ST, MA and spore inoculum level.


1986 ◽  
Vol 49 (8) ◽  
pp. 618-620 ◽  
Author(s):  
HAIM M. SOLOMON ◽  
DONALD A. KAUTTER

In October, 1983, sauteed onions in “patty-melt” sandwiches were epidemiologically responsible for a large outbreak of botulism in Peoria, Illinois. Spores of strains of Clostridium botulinum type A, recovered from Spanish onions or from patients who consumed sauteed onions, produced high toxin titers within 48 h from 2 spores/g of onions when experimentally inoculated into sauteed onions. Laboratory strains of C. botulinum type A which normally produce high-titered toxin in culture media yielded very low toxin titers and required 3 to 4 d and an extremely high inoculum of spores/g of onions. Five strains of C. botulinum type A were isolated from 75 raw onions obtained from the Peoria restaurant where the outbreak occurred.


1990 ◽  
Vol 53 (12) ◽  
pp. 1025-1032 ◽  
Author(s):  
HIROYUKI IMAI ◽  
KATSUNORI OSHITA ◽  
HIKOTAKA HASHIMOTO ◽  
DANJI FUKUSHIMA

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.


1994 ◽  
Vol 57 (10) ◽  
pp. 878-881 ◽  
Author(s):  
HAIM M. SOLOMON ◽  
E. JEFFERY RHODEHAMEL ◽  
DONALD A. KAUTTER

The ability of Clostridium botulinum type A or B spores to grow and produce toxin in fresh raw potatoes under vacuum with or without sulfite at 22°C was investigated. Fresh, peeled, sliced potatoes, untreated or dipped for 2 min in sulfite (NaHSO3) and drained, were surface-inoculated at several levels with a mixture of C. botulinum spores, either type A or B, and placed in oxygen-impermeable bags (200 g/bag) that were then vacuum-sealed and incubated at room temperature (22°C). Toxicity was tested on days 0, 3, 4, 5 and 6. After incubation, the potatoes were blended and centrifuged, and the millipore-filtered supernatant fluid was injected intraperitoneally into mice. Sensory evaluation, except taste, was also performed. Potatoes inoculated with C. botulinum type A spores, but untreated with NaHSO3 became toxic in 3 days, which coincided with the sensory evaluation, “Unfit for human consumption.” However, despite inoculum size or residual SO2 levels, potatoes treated with NaHSO3 appeared acceptable for human consumption through day 6, even though they were toxic after 4 days of incubation. Toxicity from type B spores occurred later and in fewer test samples than type A. Again, the potatoes appeared acceptable but were toxic. Thus, although NaHSO3 markedly extended the consumer acceptability of peeled, sliced, raw potatoes at the abuse temperature, it did not inhibit outgrowth and toxin production by C. botulinum under these same conditions.


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