1,720,999 research outputs found
A new scaleable method for the purification of botulinum neurotoxin type E
No full textBotulinum neurotoxins belong to the most toxic substances in nature. Well-known as a food poisoning agent for almost two centuries, the beneficial aspects of this bacterial metabolite were rediscovered about 30 years ago, These toxins, which are produced by the anaerobic bacterium Clostridium botulinum are nowadays used to treat a Variety of neuro-muscular disorders. The increased demand requires techniques for the production and purification of these toxins on an industrial scale. The method described herein is based on filtration and chromatography procedures only, Precipitation, centrifugation and dialysis steps were consequently excluded to develop a protocol, which can easily be scaled up from the laboratory purification to industrial needs. About 4 mg of BoNT/E were purified from a 10-L batch culture corresponding to in overall recovery of similar to 14%. (c) 2005 Elsevier B.V. All rights reserved
Diagnosis of botulism since 1995. Report of test results
No full textSince 1970 our laboratory is specialized in diagnosis of clostridial. diseases, including Clostridium botulinum and botulism. Since 1995, samples from more than 900 suspected botulinal cases were received, mainly in cattle, horses and men. 524 outbreaks were diagnosed as clearly positive by toxin neutralisation; 83 cases remained inconclusive with the toxin neutralisation. The geographical distribution of the positive cases in Germany is demonstrated for cattle and horses. Dispatch and treatment of specimens and interpretation of results are discussed
Determination of free DNA in soils
No full textThe concentration of free DNA in soils has to be determined in order to understand the fate and the transport of extracellular DNA, A protocol for the extraction and determination of free DNA was developed. The procedure uses separation steps, i.e. centrifugation and ultra-filtration. The free dsDNA was stained with PicoGreen(R) and determined fluorimetrically. Samples from different soils, different soil horizons, soil waters and under different land use systems were analyzed. It was found that in nearly all samples free DNA was detectable. Free DNA concentrations of up to 1950 ng (g dried sample)(-1) could be detected depending on depth, soil type and system of land use
Detection of botulinum neurotoxin - an overview and outlook
No full textLaboratory methods for the detection of botulinum toxins are insufficient as indicated in recent discussions,on bio-terrorism. Diagnostic veterinary laboratories experience problems with the absence of suitable methods with an increasing number of samples. The mouse bioassay remains the diagnostic method of choice. Alternative methods have, however, been developed, which could replace the use of animals in some instances. Data on test performance with complex sample material is only available for the mouse assay. All other assays have been tested with purified neurotoxins or culture supernatants. This short and incomplete review lists several suitable methods which are recommended for routine laboratory use
From bacterial spore to death of the patient - the development cascade of botulism
No full textThe disease >>botulism>botulinomics<< starting with Clostridium botulinum in the environment resulting in contamination of feed, followed by toxinogenesis, toxic effects and finally to clinical disease. The authors' experience of the disease over the last few years together with a review of the published literature is discussed
Diagnosis of botulism since 1995. Report of test results
No full textSince 1970 our laboratory is specialized in diagnosis of clostridial. diseases, including Clostridium botulinum and botulism. Since 1995, samples from more than 900 suspected botulinal cases were received, mainly in cattle, horses and men. 524 outbreaks were diagnosed as clearly positive by toxin neutralisation; 83 cases remained inconclusive with the toxin neutralisation. The geographical distribution of the positive cases in Germany is demonstrated for cattle and horses. Dispatch and treatment of specimens and interpretation of results are discussed
Indications on the occurrence of bovine botulism in Germany according to laboratory investigations in the years 1996-2010
No full textSince 1995 we have examined specimens for botulinum toxins and Clostridium (C.) botulinum, respectively. From our data we compiled those farms where a suspected clinical diagnosis of botulism was confirmed by a positive laboratory result (mouse bio-assay), and from which the correct postal address was given. The geographical distribution of these farms is shown in a map of Germany and compared to the concentration of cattle in the year 2007. From 1996 until 2010 1108 farms with botulinum related problems in 525 post code areas were identified. There is no correlation between cattle concentration and disease prevalence. As C. botulinum is a soil bacterium it must be presumed that also in the future this pathogen may have an influence on herd health in the affected regions
Indications on the occurrence of bovine botulism in Germany according to laboratory investigations in the years 1996-2010
No full textSince 1995 we have examined specimens for botulinum toxins and Clostridium (C.) botulinum, respectively. From our data we compiled those farms where a suspected clinical diagnosis of botulism was confirmed by a positive laboratory result (mouse bio-assay), and from which the correct postal address was given. The geographical distribution of these farms is shown in a map of Germany and compared to the concentration of cattle in the year 2007. From 1996 until 2010 1108 farms with botulinum related problems in 525 post code areas were identified. There is no correlation between cattle concentration and disease prevalence. As C. botulinum is a soil bacterium it must be presumed that also in the future this pathogen may have an influence on herd health in the affected regions
Botulinum toxins - Cause of botulism and systemic diseases?
No full textToxins of Clostridium botulinum (types A-G) are known as 'neurotoxins', causing the clinically well-known picture of flaccid muscular paralysis. The molecular biological background is the blocking of acetylcholine secretion in neuromuscular junctions by enzymatic cleavage of molecules forming the machinery of exocytosis. Two 'non-neurotoxins' (types C2, C3) are produced by some strains of C. botulinum types C and D. These affect the cytoskeleton by ribosylating actin filaments. All these toxins are used as cell biological tools for the study of specific actions and effects in different eukaryotic cells. Pharmaceutical and molecular biological research has shown their influence on several crucial organs (or cell cultures thereof) of humans and animals (brain and spinal cord, cerebellum, hippocampus, hypophysis, pancreas, adrenal glands, salivary glands and others). Under natural conditions, botulinum toxins may pass the intestinal barrier and circulate in the bloodstream for a certain time. Carriers occurring naturally in food, such as wheat germ agglutinin, digitonin or saponin, and bacterial toxins such as streptolysin O, perfringolysins, C2 toxin or botulinolysin may also form pores in cell walls. They facilitate the entry of botulinum toxins into cells that may not have natural binding receptors. It is concluded that in vivo actions of different botulinum toxins after their entry into the organism may contribute to the onset of different diseases of hitherto cryptogenic origin. Some examples are given and future problems are discussed
Persistence and mobility of a Clostridium botulinum spore population introduced to soil with spiked compost
No full textIn a recent study it could be shown that compost samples can contain Clostridium botulinum. It was investigated if C. botulinum introduced with compost into botulinum-free soil can persist and be translocated within the soil. Compost was spiked with two C. botulinum type D spore concentrations (10(3) and 10(5) spores g(-1)) and the composts were spread on an experimental site. Over a period of 939 days, samples were taken from the upper (0-5 cm) and the lower (10-30 cm) soil horizons. Physical and chemical as well as microbiological variables were measured. Clostridium botulinum spores were quantified in a culture MPN-PCR assay. On day 757 the last positive sample was obtained in the plots with the lower spore concentration (10(3) g(-1)). The bacteria were never detected in the samples taken from the lower horizons of these plots. Clostridium botulinum persisted over the whole investigation period in the plots which were treated with compost spiked with 10(5) spores g(-1). The concentrations found were between 20 and 20 000 spores g(-1) soil. The bacteria were vertically translocated and could be found in the lower soil horizons (20-2000 spores g(-1) soil) starting 70 days after the compost was spread
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