RESUMEN
Periodontological grafts are materials used in dentistry to regenerate lost gingival soft tissues or bone parts. In the case of direct contact with blood, the possibility of disease transmission from the source to the patient is high. This source can be an animal or a human. Therefore, the sterilization of grafts before implanting to the patient is of significant importance. The purpose of this study was to evaluate gamma radiation and microwave sterilization processes from microbiological and sterility perspectives and to compare the effectiveness of these two sterilization methods. Grafts were irradiated with 2, 4, 5, 10, 25 and 50 kGy doses of gamma radiation. Another group of same materials was irradiated by microwave for 1, 2, 3 and 4 min at 24,500 MHz and 900 W. Gamma radiation and microwave sterilization methods were evaluated as successful at minimum doses as 5 kGy and 3 min, respectively. Both gamma and microwave sterilization successfu lly sterilized periodontological grafts coded as PBG1, HBG1, HL1, PDG1, MBG3, MDG2 and PDG3. Moreover, microwave sterilization can be used as an alternative novel method to gamma radiation sterilization.
Asunto(s)
Bacillus pumilus/efectos de la radiación , Rayos gamma , Microondas , Esterilización/métodos , Trasplantes/efectos de la radiación , Pérdida de Hueso Alveolar/cirugía , Animales , Bacillus pumilus/crecimiento & desarrollo , Trasplante Óseo/métodos , Colágeno/efectos de la radiación , Recuento de Colonia Microbiana , Relación Dosis-Respuesta en la Radiación , Caballos , Humanos , Porcinos , Trasplantes/microbiologíaRESUMEN
Bacterial spores are one of the most resilient life forms on earth and are involved in many human diseases, such as infectious diarrhea, fatal paralytic illnesses and respiratory infections. Here, we investigated the mechanisms involved in the death of Bacillus pumilus spores after exposure to electric arcs in water. Cutting-edge microscopies at the nanoscale did not reveal any structural disorganization of spores exposed to electric arcs. This result suggested the absence of physical destruction by a propagating shock wave or an exposure to an electric field. However, Pulsed-Field Gel Electrophoresis (PFGE) revealed genomic DNA damage induced by UV radiation and Reactive Oxygen Species (ROS). UV induced single-strand DNA breaks and thymine dimers while ROS were mainly involved in base excision. Our findings revealed a correlation between DNA damage and the treatment of spores with electrical discharges.
Asunto(s)
Daño del ADN/efectos de la radiación , ADN Bacteriano/efectos de la radiación , Electricidad , Esporas Bacterianas/genética , Purificación del Agua/métodos , Bacillus pumilus/genética , Bacillus pumilus/metabolismo , Bacillus pumilus/efectos de la radiación , Infecciones Bacterianas/prevención & control , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Genoma Bacteriano/genética , Genoma Bacteriano/efectos de la radiación , Humanos , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Esporas Bacterianas/metabolismo , Esporas Bacterianas/efectos de la radiación , Rayos Ultravioleta , Microbiología del AguaRESUMEN
In this study, we investigated the inactivation efficacy of endospore-forming bacteria, Bacillus pumilus, irradiated by low-energy X-rays of different beam qualities. The different low-energy X-rays studied had cut-off energies of 50, 100 and 150 keV. Bacillus pumilus spores (in biological indicator strips) were irradiated at step doses between 6.5 to 390 Gy. The resulting bacteria populations were then quantified by a pour plate method. Results showed that X-rays of lower energies were more effective in inactivating bacterial spores. In addition, an increment in bacterial population was observed at doses below 13Gy. We attributed this increase to a radiation-induced activation of bacterial spores. Four kinetic models were then evaluated for their prediction of bacterial spore behavior under irradiation. This included: (i) first-order kinetics model; (ii) Shull model; (iii) Sapru model; and (iv) probabilistic model. From R2 and AIC analyses, we noted that the probabilistic model performed the best, followed by the Sapru model. We highlighted that for simplicity in curve fitting the Sapru model should be used instead of the probabilistic model. A 12-log reduction in bacterial population (corresponding to a sterility assurance level of 10-6 as required in the sterilization of medical devices) was computed to be achievable at doses of 1000, 1600 and 2300 Gy for the three different X-ray cut-off energies respectively. These doses are an order in magnitude lesser than that required in gamma irradiation. This highlights the applicability of cheaper and safer table-top X-ray sources for sterilization application.
Asunto(s)
Bacillus pumilus/efectos de la radiación , Viabilidad Microbiana/efectos de la radiación , Esporas Bacterianas/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Cinética , Modelos Teóricos , Rayos XRESUMEN
In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be ß-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain.