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Shelf-stable canned butter is currently available in retail stores, and many home-preservationists promote home-canning of butter. Non-cultured butter is a low-acid canned food, which would presumably require thermal processing. The lack of a thermal process step in canned butter products raises questions of potential safety, because they are hermetically sealed and generally exhibit anaerobic growth conditions, which are optimal for Clostridium botulinum growth. Without thermal processing, low-acid canned foods (LACF) must have inhibitory factors present to prevent C. botulinum growth. Some potential intrinsic inhibitory factors, or "hurdles", within butter include: reduced water activity (aw), acidity (pH) in cultured products, elevated salt content, and the micro-droplet nature of the aqueous phase in the butter emulsion. It was hypothesized that a normal intact butter emulsion would have sufficient "hurdles" to prevent C. botulinum growth, while a broken butter emulsion would result in a larger aqueous phase that would allow for growth. Butter was prepared using a batch churn method with either inoculated or uninoculated cream. Butter samples with four different salt amounts (0, 0.8, 1.6, & 2.4% added NaCl) were prepared and placed in coated aluminum cans for storage. Samples were stored for 1 or 2 week periods at either 22°C or 41°C and then plated for C. sporogenes growth. Samples stored at 41°C showed a significant increase over those stored at 22°C. This growth increase occurred due to incubation near the optimal growth temperature for C. sporogenes and damage to emulsion structure. Furthermore, sodium chloride (NaCl) addition was found to have a significant effect on C. sporogenes growth, with 0.8 % NaCl promoting more growth than 0%, but with decreases in growth beyond 0.8%. Uninoculated control plates were also found to have bacterial growth. This growth was attributed to other anaerobic bacteria present within the cream. It was concluded that removal of the butter structure "hurdle" could result in C. botulinum growth even at elevated salt levels and therefore home preparation of canned butter is not advisable. It is also possible that commercially canned butter, if heat abused, could potentially allow for C. botulinum growth and therefore consumption is not recommended.

A procedure to rapidly screen Clostridium botulinum type A and Clostridium sporogenes for plasmids was developed. Plasmid profiles of five C. botulinum type A strains and seven C. sporogenes strains were determined and a possible relationship of plasmids to toxin production was examined. The differentiation of these organisms by plasmid fingerprinting was also studied. The plasmid isolation procedure was a modified cleared lysate technique, including lysis under alkaline conditions. Samples were subject to agarose gel electrophoresis to detect plasmid DNA. Culture age affected plasmid detection due to changes in the cell density and lysing efficiency. Middle to late log cultures were used throughout the study because they provided optimum plasmid detection. Four out of five C. botulinum type A strains and three out of seven C. sporogenes strains contained extrachromosomal DNA. For those C. botulinum type A strains which contained plasmids, there were always two, one 15 to 15.6 Mdal and the other 6.2 Mdal. C. sporogenes showed less consistency in plasmid size and number and their plasmids were generally of a greater molecular weight than those in C. botulinum type A. One C. sporogenes strain contained two plasmids and two strains contained one plasmid. Toxin production may be plasmid-mediated in the plasmid containing strains, but there was no apparent general relationship, because one of the toxic strains did not show the presence of plasmids. Plasmid screening may be useful in the differentiation of these closely related organisms without toxin testing. / Master of Science

Benign masseter hypertrophy causes swelling at the angulus mandibulae and may be associated with masticatory myofascial pain due to hyperfunction from bruxism. The aim of this research was to use the systematic review process to investigate the true or reliable scientific evidence contained in four major databases pertaining to the efficacy and safety of intra-muscular injections of botulinum toxin A (BTX-A) for the treatment of masticatory myofascial pain and benign masseter hypertrophy associated with bruxism, compared with placebo or other traditional treatments prescribed for bruxism such as occlusal splints, pharmacotherapy, or lifestyle modification. Using the PICO format, a research question was formulated, MeSH terms were derived, and an electronic literature search was conducted in PubMed, Embase, Web of Science, and Cochrane. This sequence was followed by a screening and selection of articles by two independent reviewers according to defined inclusion and exclusion criteria. The selected studies were then evaluated and assessed based on study quality and identification of biases, and the results were summarized and reported. This review highlighted the lack of well-designed, randomized controlled trials to evaluate the efficacy and safety of botulinum toxin A for reducing the size/volume of the masseter muscles and for improving masticatory myofascial pain in patients who present with bruxism. Thus, the results were inconclusive.

The risk of toxin production by Clostridium botulinum type E in pasteurized oysters was evaluated. Thermal death time studies for type E spores in a oyster homogenate showed that the spores survived pasteurization at 550 C; D-values ranged from 65 to 100 min at 700 C and 880 to 1300 min at 550 C. When tubes of oyster homogenate were inoculated with 1x103 type E spores/ml and stored at 3, 6, 12, and 30oC, no growth was observed; however, when the inocculum was increased to 1x104 spores/ml, toxin was produced during storage at 30oC, but not at lower temperatures. For containers of whole oysters inoculated with a low spore level, toxin production occurred during incubation at 6, 12, and 30oC. / Master of Science

Toxins produced by the Gram-positive bacteria Clostridium botulinum cause transient chemodenervation of mammalian muscle. The toxin binds to specific proteins within cholinergic presynaptic nerve terminals which regulate the release of acetylcholine in the synaptic space resulting is loss of muscle activation and function. Local injections with botulinum toxins are currently used in humans for the treatment of disorders that benefit from prolonged neuromuscular blockade such as strabismus, blepharospasm, focal dystonias, spasticity, tremors, and anal fissures. Injections with botulinum toxin type A into the internal or external anal sphincter cause relaxation of the anal canal and allow healing of chronic anal fissures. Perineal lacerations in mares, which occur during foaling often dehisce after surgical repair due to the high pressure across the incision resulting from accumulation of feces in the rectum. We hypothesized local injections of Clostridium botulinum type B toxin into the external anal sphincter could cause a decrease in anal pressures, thus reducing the incidence of dehiscence if used before surgical repair of perineal laceration in mares. The purpose of this project was to determine the effects of BTB injection in the external anal sphincter in normal horses. Our hypothesis was that local injection of BTB would result in transient reduction of anal tone without causing clinical side effects. Peak and resting anal sphincter pressures of horses were measured with a custom made rectal probe connected to a pressure transducer. Pressures were measured before treatment and after injection with Clostridium botulinum type B toxin (BTB) or saline. Dose titration with 500, 1000, 1500 and 2500 units of BTB was completed. The horses' physical changes, behavior, and anal pressure were recorded. Injection of 1000 units of BTB produced significant reduction in peak anal pressure from days 2 to 84 when compared to control animals (P<0.05). Maximal effect of the toxin was observed within the first 15 days after injections followed by a slow return to baseline over 168 days. Injection in the anal sphincter with 2500 units of BTB in one horse produced signs of depression, generalized weakness, and dysphagia for 14 days. Clinical side effects were not observed in horses after injections with 500, 1000, or 1500 units of BTB. In summary, local injections of botulinum toxin type-B in the external anal sphincter of horses caused transient relaxation of the anus and reduction of peak anal pressures. Systemic side effects were observed in one horse, which suggested a narrow dosage range to avoid toxicity. Further research to test the effects of botulinum toxin in clinical cases is needed to determine the full potential of this treatment modality. / Master of Science

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Syfte Syftet med föreliggande litteraturstudie är att jämföra behandlingseffekten av kliniska studier som utgår från patienter med käkledssjukdomar, temporomandibulär dysfunktion, TMD och bruxism  där behandlingsalternativen stabiliseringsskena och/eller läkemedlet botulinumtoxin (botox) inkluderats.    Material och metod Litteraturstudien genomfördes via PICO-systemet, med litteratursökning i databaserna Medline/PubMed, Web of Science och Scopus. Insamlad data bearbetades genom en granskningsprocess på titel-, sammanfattnings- och slutligen fulltextnivå, utifrån förutbestämda inklusions- och exklusionskriterier.   Resultat Två studier uppfyllde urvalskriterierna. Behandlingsterapin med botox och stabiliseringsskena kunde följas under sex månaders tid i båda studierna. Ena studien bestod av 40 patienter som delades in i två grupper, en för varje behandlingsmetod med fyra uppföljnings- och behandlingstillfällen. Den andra studien bestod av 73 patienter som delades in i tre grupper, en för respektive behandlingsmetod och en grupp som inkluderade båda behandlingsmetoderna, med tre uppföljningstillfällen. Antal enheter och placering av botoxinjektioner skilde sig åt i studierna, tillika antal timmars användning av stabiliseringsskena. Studierna presenterade trots skillnader liknande resultat.    Slutsats  Behandling med stabiliseringsskena är inte nödvändig för patienter som behandlats med botulinumtoxin, eftersom de statistiska resultaten inte påvisade någon större skillnad i effekt. Behandling med botulinumtoxin och dess effekt på patientens besvär beror på antal enheter av läkemedlet och placering vid injektion samt i jämförelse med omfattningen av hur mycket patienten använder stabiliseringsskenan. Botulinumtoxin kan användas som behandlingsmetod när patienten av olika anledningar inte kan använda stabiliseringsskena. / Purpose The purpose of this literature study is to compare clinical studies that have included the treatment options of using stabilization appliances and/or medication with botulinum toxin. The included studies must have included and treated patients with temporomandibular disorders, temporomandibular dysfunction, TMD and bruxism.    Material and method The literature study was executed based on the PICO system and through searches in the databases Medline/PubMed, Web of Science and Scopus. Subsequently, a review process was carried out at title, abstract and finally full text level, based on predetermined inclusion and exclusion criteria.   Results Two studies met the selection criteria. The treatment therapy could be followed for a period of six months in both studies. One study consisted of 40 patients who were divided into two groups, one for each treatment method with four follow-up and treatment sessions. The second study consisted of 73 patients who were divided into three groups, one for each treatment method and one group that included both treatment methods and three follow-up sessions. Units and location of botulinum toxin injections and the hours of use of the stabilization appliance differed slightly in the two studies. However, the studies reported similar outcomes.   Conclusion The treatment with stabilization appliance is not necessary for patients who are being treated with botulinum toxin due to non-statistical significance reported. The outcome of botulinum toxin treatment may depend on the location and number of units of the medication in comparision to the extent to which the patient uses the stabilization appliance. Botulinum toxin may gain as a treatment method when the patient is unable to use a stabilization appliance for various reasons.

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