Detection Of Clostridium Botulinum Research Articles

Detection of Clostridium botulinum in Some Egyptian Fish Products, Its Control In Vitro Using Citrus Leaves Extracts, and Applicability of Citrus limon Leaf Extract in Tuna

This study aims to detect Clostridium botulinum and its control using natural leaf extracts of Citrus limon, Citrus sinensis, and Citrus unshiu in Egyptian fish products, e.g., canned tuna, canned sardine, canned mackerel, fesikh, moloha, and renga, as well the application of C. limon in tuna. Moreover, the antibacterial activity of the C. limon leaf extract was also estimated. In the water extract, ascorbic acid, total flavonoid content (TFC), and total phenolic content (TPC) were determined by volumetric, aluminum chloride, and Folin-Ciocalteu approaches, respectively. The antioxidant ability of the extract was analyzed in vitro via free radical scavenging (DPPH) and Ferric reducing assays. The results showed variability in the distribution of the total number of positive C. botulinum in fish samples from three different governorates under study, which were (24) Alexandria, (16) Beheira, and (17) Gharbia, out of the 120 tested samples in each governorate. Additionally, the findings revealed that all three Citrus extracts contain an appropriate number of secondary metabolites, with a sustainable presence of saponin and tannins in the C. limon extract. Furthermore, all Citrus extracts inhibited bacterial growth by increasing the inhibition zone, with C. limon being the best extract (25 mm) compared to C. sinensis and C. unshiu. The overall results showed the high antioxidant and anti-Clostridium powers (p < 0.05) of C. limon leaf extract, indicating its preservative activity in fishery products during storage. Finally, C. limon leaf extract can fight off C. botulinum and is considered a promising natural preservation candidate in ensuring safe and fresh fishery products.

Detection of Clostridium botulinum in liquid manure and biogas plant wastes.

Biogas plants have been considered as a source for possible amplification and distribution of pathogenic bacteria capable of causing severe infections in humans and animals. Manure and biogas wastes could be sources for spore-forming bacteria such as Clostridium botulinum. In the present study, 24 liquid manure and 84 biogas waste samples from dairies where the majority of the cows suffered from chronic botulism were investigated for the presence of botulinum neurotoxins (BoNT) and C. botulinum spores. The prevalence of BoNT/A, B, C, D, and E in biogas wastes was 16.6, 8.3, 10.7, 7.1, and 10.8 %, respectively, while in manure, the prevalence was 0.0, 0.0, 0.0, 8.3, and 4.1 %, respectively. After enrichment of samples in reinforced cultural medium, they were tested for C. botulinum BoNT/A, B, C, D, and E using ELISA (indirect C. botulinum detection). The prevalence of C. botulinum type A, B, C, D, and E samples in biogas wastes was 20.2, 15.5, 19, 10.7, and 34.8 %, respectively, while the prevalence in liquid manure was 0.0, 0.0, 0.0, 8.3, and 12.5 %, respectively. In conclusion, the occurrence of BoNT and C. botulinum spores in biogas waste of diseased animals indicates an increased and underestimated hygienic risk. Application of digestates from biogas fermentations as fertilizers could lead to an accumulation of long lifespan spores in the environment and could be a possible health hazard.

Methods and difficulties in detection of Clostridium botulinum and its toxins.

The aim of this work was to present selected data regarding traditional and modern methods for C. botulinum and its toxins detection. In this article, methods based on culturing techniques, mouse bioassay, immunological techniques, chromatography and PCR, PFGE, RFLP, AFLP are described. The mentioned techniques were evaluated considering their usefulness in the samples examination, genotyping of strains and the diagnostics of botulism.

Detection of Clostridium botulinum in fish and environmental samples using polymerase chain reaction

A test protocol for the detection and enumeration of Clostridium botulinum in fish and sediment samples with specific identification of neurotoxin types A, B, E and F was developed. Specific amplification products generated by polymerase chain reaction (PCR) formed the basis of identification of the toxin-producing organism, whereas quantification of the results was achieved with the most probable number (MPN) method. Twenty-six C. botulinum strains studied with PCR assays after enrichment in trypticase-peptone-glucoseyeast extract (TPGY) broth gave identical results as with the mouse bioassay. The suitability of the detection method for food and environmental surveys was assessed by running it on 32 samples of rainbow trout inoculated with spore loads ranging from 10 2 to 10 6 C. botulinum type E spores per kg. The organism was detected in all samples, and MPN estimates corresponded well to inoculum levels. In order to assess possible natural contamination, 16 fish and 16 visceral samples of rainbow trout, as well as ten aquatic sediment samples were tested. Of these, eight (80%) of the sediment samples were positive, with estimated spore counts of C. botulinum type E ranging from 95–2710 per kg sample.

Detection of Clostridium Botulinum in Honey by a Procedure Involving Membrane Filtration

A new procedure for the detection of botulinal spores in honey was proposed. This involved dilution of honey samples, heating for spore activation and for accelerated filtration, membrane filtration, incubation of the membranes in a recovery medium, and toxin assays. The numbers of spores of Clostridium botulinum types A and B added and the numbers of spores recovered by this method were essentially the same.

An Improved Cooked Meat Medium for the Detection of Clostridium botulinum

A new medium composed of cooked meat and fluid thioglycolate broth was tested with 20 proteolytic and 11 saccharolytic strains of Clostridium botulinum. All of the proteolytic strains produced a black ring at the surface of the broth, presumably due to hydrogen sulfide production, while the saccharolytic strains produced white opaque rings. Since the black ring is formed rapidly and is easily seen, the new medium may be useful for the rapid identification of proteolysis in unknown isolates. Growth and toxin production were better in the new medium than in the usual cooked meat medium or in cooked meat medium plus 0.5% glucose. These results suggest that the new medium will be useful in facilitating identification of C. botulinum.

Detection of Clostridium botulinum and Its Toxin in Food. I. Detection of Clostridium botulinum Type E in Smoked Fish

Abstract One mg trypsin/ml is incorporated into an enrichment medium to improve the detection of Clostridium botulinum type E. The procedure was evaluated using smoked whitefish chubs as well as other kinds of smoked fish inoculated with small numbers of spores of C. botulinum types A, B, and E. A greater number of positives was obtained for all smoked fish inoculated with type E spores when this medium was used for enrichment. In addition, by the use of this medium at 26°C for 5 days, proteolytic C. botulinum types A and B can also be detected. Collaborative study is recommended.

IMMUNOFLUORESCENT IDENTIFICATION OF CLOSTRIDIUM BOTULINUM.

THE detection of Clostridium botulinum usually involves culturing the suspect sample, and then testing the culture for the presence of botulinum toxin. The toxin is identified by mouse protection tests using specific botulinum antitoxins1, a procedure complicated by the known existence of six different types of Cl. botulinum2, each with its specific toxin. These culture and mouse inoculation procedures require much time and special animal facilities, and greatly hinder experimental work with Cl. botulinum.

The detection of clostridium botulinum by means of luminescent antibodies

The author obtained a conjugate of botulinus rabbit antiserum with fluorescein isocyanate, possessing immunological specificity. In staining the films of pure cultures ofCl. botulinum by the luminescent antiserum, an intensive green luminescence of these microorganisms was obtained. When preparations of other anaerobic cultures,E. coli, staphylococci or anthracoids were stained with the same conjugate, the specific luminescence was absent. In this way it is possible to reveal solitaryCl.botulinum cells in the films prepared from suspensions with a concentration of up to 500,000 bacterine cells per 1 ml. The method of staining by luminescent antibodies makes it possible to identifyCl. botulinum in microscopic preparations.