scholarly journals Honey sold directly by producers in the Silesian region of Poland as a source of Clostridium botulinum types A, B, E and F

2017 ◽  
Vol 35 (No. 3) ◽  
pp. 194-199 ◽  
Author(s):  
Fatma Hayıt ◽  
Hülya Gül
Keyword(s):  
Botulinum Toxin ◽  
Multiplex Pcr ◽  
Clostridium Botulinum ◽  
Type A ◽  
Honey Sample ◽  
Type B ◽  
Subsequent Processing ◽  
Pcr Method ◽  
Centrifugation Method ◽  
Cooked Meat

The level of contamination of honey with Clostridium botulinum spores is considered as an indicator of the adequacy of hygienic practices during collection, extraction, and subsequent processing. A total of 39 honey samples purchased directly from beekeepers at outdoor markets and from small amateur apiaries in Silesia were analysed for Clostridium botulinum spores. The samples were prepared using a dilution centrifugation method and cultured in parallel in cooked meat medium (CMM) and tripticase peptone glucose yeast (TPGY) enrichment broths. Identification of C. botulinum toxin types A, B, E, and F was performed with the use of a multiplex PCR method. The analysis showed six (15.4%) samples to be contaminated with C. botulinum spores. The major serotypes detected were type A – in two (5.1%) and type B – in two (5.1%) honey samples, respectively. Types E and F were found in 1 (2.6%) and 1 (2.6%) positive honey sample analysed, respectively.

scholarly journals Clostridium botulinum Spores Found in Honey from Small Apiaries in Poland

2016 ◽  
Vol 60 (2) ◽  
pp. 89-100 ◽  
Author(s):  
Joanna Wojtacka ◽  
Beata Wysok ◽  
Zbigniew Lipiński ◽  
Małgorzata Gomółka-Pawlicka ◽  
Helena Rybak-Chmielewska ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Multiplex Pcr ◽  
Clostridium Botulinum ◽  
Type A ◽  
Type B ◽  
Average Quantity ◽  
Pcr Method ◽  
Centrifugation Method ◽  
Cooked Meat

Abstract A total of 102 honey samples collected from small apiaries (≤ 20 hives) in Poland were analysed for the presence of Clostridium botulinum spores. The samples were prepared using the dilution centrifugation method and cultured in parallel in cooked meat medium (CMM) and tripticase peptone glucose yeast (TPGY) enrichment broths. Identification of toxin types A, B, and E of Clostridium botulinum strains was performed with the use of the multiplex PCR method. Positive samples were also subjected to quantitative analysis with the use of Clostridium botulinum Isolation Agar Base (CBAB). The prevalence analysis showed 22 (21.6%) samples contaminated with C. botulinum spores. The major serotype detected was botulin neurotoxin type A – 16 (72.7%) whereas type B was found in 3 (13.6%) honey samples and type E also only in 3 (13.6%) honey samples. Dual-toxin-producing strains were noted. The average quantity of spores in PCR - C. botulinum positive samples was 190 in 1 gram of honey.

Failure of Clostridium botulinum to Grow in Fresh Mushrooms Packaged in Plastic Film Overwraps with Holes

1978 ◽  
Vol 41 (5) ◽  
pp. 348-350 ◽  
Author(s):  
H. SUGIYAMA ◽  
KANDEE S. RUTLEDGE
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Type A ◽  
Plastic Film ◽  
Type B ◽  
Chloride Film

Fresh, commercially grown mushrooms were inoculated in the stem with Clostridium botulinum spores (2.5 × 104 of each of four type A with l × 105 of each of four type B strains). One lb (454 g) of mush-rooms. including two spore-inoculated ones. was packaged in paper-borad trays and overwrapped with a ploybinyl chloride film. The packages were incubated 6 days at 24–26 C after making one or two holes of 1/8 inch (3.175 mm) diameter in the overwrap of test packages. Botulinum toxin. either type A only or mixed with type B, was found in the spore-inoculated mushrooms of all 28 control packages (no hole), but was not detected in any from the 123 packages with one hole and the 4 7 with two holes. The O2% inside the packages after incubation averaged 1.5 for the controls. 4.0 for packages with one hole and 6.2 for the 2-hole package group.

scholarly journals Multiplex PCR Assay for Detection and Identification of Clostridium botulinum Types A, B, E, and F in Food and Fecal Material

2001 ◽  
Vol 67 (12) ◽  
pp. 5694-5699 ◽  
Author(s):  
Miia Lindström ◽  
Riikka Keto ◽  
Annukka Markkula ◽  
Mari Nevas ◽  
Sebastian Hielm ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Clostridium Botulinum ◽  
Botulinum Neurotoxin ◽  
Bacterial Species ◽  
Type A ◽  
Food Samples ◽  
Type B ◽  
Fecal Material ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

ABSTRACT Botulism is diagnosed by detecting botulinum neurotoxin andClostridium botulinum cells in the patient and in suspected food samples. In this study, a multiplex PCR assay for the detection of Clostridium botulinum types A, B, E, and F in food and fecal material was developed. The method employs four new primer pairs with equal melting temperatures, each being specific to botulinum neurotoxin gene type A, B, E, or F, and enables a simultaneous detection of the four serotypes. A total of 43 C. botulinum strains and 18 strains of other bacterial species were tested. DNA amplification fragments of 782 bp for C. botulinum type A alone, 205 bp for type B alone, 389 bp for type E alone, and 543 bp for type F alone were obtained. Other bacterial species, including C. sporogenes and the nontoxigenic nonproteolytic C. botulinum-like organisms, did not yield a PCR product. Sensitivity of the PCR for types A, E, and F was 102 cells and for type B was 10 cells per reaction mixture. With a two-step enrichment, the detection limit in food and fecal samples varied from 10−2 spore/g for types A, B, and F to 10−1 spore/g of sample material for type E. Of 72 natural food samples investigated, two were shown to contain C. botulinum type A, two contained type B, and one contained type E. The assay is sensitive and specific and provides a marked improvement in the PCR diagnostics of C. botulinum.

Incidence and Origin of Clostridium botulinum Spores in Honey

1981 ◽  
Vol 44 (11) ◽  
pp. 812-814 ◽  
Author(s):  
C. N. HUBTANEN ◽  
D. KNOX ◽  
H. SBIMANUKI
Keyword(s):  
New Jersey ◽  
Apis Mellifera ◽  
Clostridium Botulinum ◽  
Water Solution ◽  
Type A ◽  
Type B ◽  
Sugar Water ◽  
Quantitative Estimates ◽  
Foreign Countries ◽  
Cooked Meat

Eighty honey samples, including some from foreign countries, were obtained from a local processor or from apiaries in Pennsylvania, Illinois and New Jersey. They were analyzed for Clostridium botulinum spores by a dilution-centrifugation (DC) procedure and by direct addition (DA) of honey to two different enrichment media. All were negative by the DC method; five were positive by DA in fluid thioglycollate media and six by DA in cooked meat media. Some samples positive in fluid thioglycollate media were negative in cooked meat media and vice versa. Bees (Apis mellifera, 25,000 per hive) were experimentally inoculated with spores of C. botulinum by feeding a 50% sugar-water solution containing 1.6 × 105 spores of 20 strains (equal numbers of 11 type A and 9 type B). Honey collected from the hive 2 weeks later contained 1100 spores per g; that collected after 5 weeks contained 50 spores per g. Quantitative estimates of honey yield and spore contents indicated that all the spores originally ingested by the bees had been incorporated into the honey. No botulinal spores were found in the intestinal or rectal contents of the bees 2 weeks or more after spore ingestion.

In vitro and in vivo Assessment of Silver Nanoparticles Against Clostridium botulinum Type A Botulinum

2019 ◽  
Vol 16 (1) ◽  
pp. 113-119 ◽  
Author(s):  
Mohammad Aminianfar ◽  
Siavash Parvardeh ◽  
Mohsen Soleimani
Keyword(s):  
Silver Nanoparticles ◽  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Lethal Dose ◽  
Type A ◽  
Positive Control ◽  
Negative Control ◽  
Laboratory Animals ◽  
Control Group ◽  
Nano Silver

Background: Clostridium botulinum causes botulism, a serious paralytic illness that results from the ingestion of a botulinum toxin. Because silver nanoparticle products exhibit strong antimicrobial activity, applications for silver nanoparticles in healthcare have expanded. Therefore, the objective of the current study was to assess a therapeutic strategy for the treatment of botulism toxicity using silver nanoparticles. Methods: A preliminary test was conducted using doses that produce illness in laboratory animals to determine the absolute lethal dose (LD100) of botulinum toxin type A (BoNT/A) in mice. Next, the test animals were divided into six groups containing six mice each. Groups I, II and III were the negative control (botulinum toxin only), positive control-1 (nano-silver only) and positive control-2 (no treatment), respectively. The remaining groups were allocated to the toxin that was supplemented with three nano-silver treatments. Results: The mortality rates of mice caused by BoNT/A significantly reduced in the treatment groups with different doses and injection intervals of nano-silver when compared to the negative control group. BoNT/A toxicity induced by intraperitoneal injection of the toxin of Clostridium botulinum causes rapid death while when coupled with nano-osilver results in delayed death in mice. Conclusion: These results, while open to future improvement, represent a preliminary step towards the satisfactory control of BoNT/A with the use of silver nanoparticles for human protection against this bioterrorism threat. Further study in this area can elucidate the underlying mechanism for detoxifying BoNT/A by silver nanoparticles.

Production of Botulinum Toxin in Inoculated Pack Studies of Foil-Wrapped Baked Potatoes

1981 ◽  
Vol 44 (12) ◽  
pp. 896-898 ◽  
Author(s):  
H. SUGIYAMA ◽  
MARGY WOODBURN ◽  
K. H. YANG ◽  
COLLEEN MOVROYDIS
Keyword(s):  
Botulinum Toxin ◽  
Clostridium Botulinum ◽  
Aluminum Foil ◽  
Type A ◽  
Russet Burbank ◽  
Inoculation Site ◽  
Botulinum Type ◽  
Lethal Doses ◽  
Spore Inoculation

Idaho Russet Burbank potatoes were surface or stab inoculated with 10 to 105 spores of Clostridium botulinum type A strain, overwrapped in aluminum foil, baked at 204 C for 50 min or 96 C for 3 h and then held at 22 or 30 C. The shortest incubations resulting in the first botulinogenic potatoes were inversely related to spore doses and ranged from 3 to 7 days; potatoes inoculated with 10 spores were toxic after 5 to 7 days. Total toxin in individual potatoes incubated 3 to 5 days were 5 × 103 to 5 × 105 mouse mean lethal doses. Toxin was not found at distances greater than 1.6 cm from the spore inoculation site. Results indicate that left-over, foil-wrapped, baked potatoes are a perishable food that must be refrigerated.

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