Sporulation and Toxin Production by Clostridium botulinum Type G

1979 ◽  
Vol 42 (12) ◽  
pp. 965-967 ◽  
Author(s):  
H. M. SOLOMON ◽  
D. A. KAUTTER
Keyword(s):  
Comparative Study ◽  
Intraperitoneal Injection ◽  
Clostridium Botulinum ◽  
Solid Medium ◽  
Toxin Production ◽  
Optimum Conditions ◽  
Liquid Media ◽  
Botulinum Type ◽  
The Media

A comparative study was conducted to determine the optimum conditions for sporulation and toxin production by Clostridium botulinum type G, strain 89. One solid and four liquid media were compared for their ability to promote sporulation. After being inoculated, the media were incubated at 35 C for 12 days, at 30 C for 16 days, and at 26 C for 21 days. Spores were harvested by centrifugation, washed 3 times and resuspended to give a 35 × concentration, then counted by the MPN procedure. Spores grown on the solid medium at 35 C for 16 days gave higher counts than those grown at the same temperature in the liquid media. Toxin production was studied in eight media at 35, 30 and 26 C over a 24-day period with samplings every 2 to 3 days. Three of the media contained trypsin and five were trypsinized after growth. Toxin titers were determined by intraperitoneal injection of mice and expressed as MLD/ml of culture. Higher toxin titers were obtained at 26 and 30 C in media containing 0.4% glucose.

Inhibition of Clostridium botulinum Growth and Toxigenesis in a Model Gravy System by Coinoculation With Bacteriocin-Producing Lactic Acid Bacteria

1993 ◽  
Vol 56 (6) ◽  
pp. 485-488 ◽  
Author(s):  
ALLISON D. CRANDALL ◽  
THOMAS J. MONTVILLE
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Acid Production ◽  
Clostridium Botulinum ◽  
Toxin Production ◽  
Bacteriocin Production ◽  
Botulinum Type ◽  
The Media ◽  
Mild Temperature

The ability of several lactic acid bacteria (LAB) to inhibit Clostridium botulinum toxigenesis was investigated. Acidification studies identified the bacteriocinogenic strains Lactococcus lactis ATCC 11454 and Pediococcus pentosaceus ATCC 43200 as the most promising based on their ability to rapidly acidify a model gravy system. These two strains, a third bacteriocinogenic strain Lactobacillus plantarum BN, and nonbacteriocinogenic strains as controls were then coinoculated along with C. botulinum type A and B spores into a model gravy system to determine if bacteriocin production and acidification are effective in preventing C. botulinum growth and toxin production. Triplicate tubes of gravy-like media containing either 0 or 0.5% glucose were coinoculated with the LAB at 104 CFU/ml and with the pool of heat-shocked C. botulinum spores at 102, 104, and 106 CFU/ml and incubated anaerobically at 15, 25, or 35°C. The media were monitored for C. botulinum growth, toxin production, and acidity. At 15°C, both the bacteriocinogenic and nonbacteriocinogenic strains of L. lactis and L. plantarum prevented toxigenesis in gravy containing glucose at all C. botulinum inocula levels. The bacteriocinogenic and nonbacteriocinogenic strains of P. pentosaceus prevented toxin production by C. botulinum at 102 and 104 CFU/ml in the presence of glucose. P. pentosaceus 43200 was the only strain tested showing inhibition in the absence of glucose, preventing toxigenesis by C. botulinum at 102 CFU/ml. At 25 and 35°C, none of the lactic acid bacteria tested prevented toxigenesis. The results suggest that acid production by these strains of LAB may afford some protection against mild temperature abuse and that bacteriocin production had little if any additional effect. The biopreservation system was ineffective at temperatures of 25 and 35°C.

Effect of Salt Level and Nitrite on Toxin Production by Clostridium botulinum Type E Spores in Smoked Great Lakes Whitefish1

1987 ◽  
Vol 50 (3) ◽  
pp. 212-217 ◽  
Author(s):  
S. L. CUPPETT ◽  
J. I. GRAY ◽  
J. J. PESTKA ◽  
A. M. BOOREN ◽  
J. F. PRICE ◽  
...  
Keyword(s):  
Great Lakes ◽  
Water Activity ◽  
Salt Concentration ◽  
Clostridium Botulinum ◽  
Toxin Production ◽  
Water Phase ◽  
Salt Level ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E ◽  
Average Water

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.

Toxin Production by Clostridium botulinum Type E in Packaged Channel Catfish

1997 ◽  
Vol 60 (11) ◽  
pp. 1358-1363 ◽  
Author(s):  
PING CAI ◽  
MARK A. HARRISON ◽  
YAO-WEN HUANG ◽  
JUAN L. SILVA
Keyword(s):  
Channel Catfish ◽  
Clostridium Botulinum ◽  
Modified Atmosphere Packaging ◽  
Toxin Production ◽  
Storage Conditions ◽  
Mouse Bioassay ◽  
Modified Atmosphere ◽  
Oxygen Barrier ◽  
Botulinum Type ◽  
Clostridium Botulinum Type E

Channel catfish were inoculated with 3 to 4 log spores/g of a mixed pool of four strains of C. botulinum type E (Beluga, Minnesota, G21-5, and 070) and were packaged with an oxygen-permeable overwrap, in an oxygen-barrier bag with a modified atmosphere of CO2-N2 (80:20) or in a master bag with the same modified atmosphere. Packaged fish were stored at either 4°C and sampled at intervals over 30 days or at 10°C and sampled at intervals over 12 days. An additional master bag treatment in which overwrap-packaged catfish was stored first at 4°C, then removed from the master bags and stored at 10°C, was sampled at intervals over 18 days. Toxin production was evaluated using the mouse bioassay. Aerobic psychrotrophic and anaerobic populations were enumerated, and product spoilage characteristics were noted. Under abusive storage conditions of 10°C, there was no difference among the potential for toxin production in the packaged fish, with botulinum toxin detected on fish from each package type by day 6. At 4°C, toxin production was detected on day 9 in the overwrapped packages, while it was on day 18 in the modified atmosphere packaging. No toxin was found in the master bags held continually at 4°C. Toxin was detected on day 18 from samples initially held at 4°C in the master bag and subsequently held at 10°C. Spoilage preceded toxin production for samples stored at 4°C for each type of packaging. At 10°C, spoilage and toxin detection times coincided.

Risk of Growth and Toxin Production by Clostridium botulinum Nonproteolytic Types B, E, and F in Salmon Fillets Stored Under Modified Atmospheres at Low and Abused Temperatures

1987 ◽  
Vol 50 (4) ◽  
pp. 330-336 ◽  
Author(s):  
GENERO W. GARCIA ◽  
CONSTANTIN GENIGEORGIS ◽  
SEPPO LINDROTH
Keyword(s):  
Clostridium Botulinum ◽  
Storage Time ◽  
Toxin Production ◽  
Lag Phase ◽  
Type B ◽  
Inoculum Level ◽  
Modified Atmospheres ◽  
Botulinum Type ◽  
Best Fit ◽  
Design Experiments

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.

Growth and Toxin Production by Clostridium botulinum in Sauteed Onions

1986 ◽  
Vol 49 (8) ◽  
pp. 618-620 ◽  
Author(s):  
HAIM M. SOLOMON ◽  
DONALD A. KAUTTER
Keyword(s):  
Clostridium Botulinum ◽  
Culture Media ◽  
Type A ◽  
Toxin Production ◽  
High Inoculum ◽  
Botulinum Type ◽  
Large Outbreak

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.

Growth and Toxin Production by Clostridium botulinum in Sliced Raw Potatoes Under Vacuum With and Without Sulfite

1994 ◽  
Vol 57 (10) ◽  
pp. 878-881 ◽  
Author(s):  
HAIM M. SOLOMON ◽  
E. JEFFERY RHODEHAMEL ◽  
DONALD A. KAUTTER
Keyword(s):  
Sensory Evaluation ◽  
Clostridium Botulinum ◽  
Room Temperature ◽  
Type A ◽  
Toxin Production ◽  
Inoculum Size ◽  
Consumer Acceptability ◽  
Human Consumption ◽  
Type B ◽  
Botulinum Type

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.

Sign in / Sign up

Export Citation Format

Share Document