BackgroundGluconacetobacter diazotrophicus Pal5 is an endophytic diazotrophic bacterium that lives in association with sugarcane plants. It has important biotechnological features such as nitrogen fixation, plant growth promotion, sugar metabolism pathways, secretion of organic acids, synthesis of auxin and the occurrence of bacteriocins.ResultsGluconacetobacter diazotrophicus Pal5 is the third diazotrophic endophytic bacterium to be completely sequenced. Its genome is composed of a 3.9 Mb chromosome and 2 plasmids of 16.6 and 38.8 kb, respectively. We annotated 3,938 coding sequences which reveal several characteristics related to the endophytic lifestyle such as nitrogen fixation, plant growth promotion, sugar metabolism, transport systems, synthesis of auxin and the occurrence of bacteriocins. Genomic analysis identified a core component of 894 genes shared with phylogenetically related bacteria. Gene clusters for gum-like polysaccharide biosynthesis, tad pilus, quorum sensing, for modulation of plant growth by indole acetic acid and mechanisms involved in tolerance to acidic conditions were identified and may be related to the sugarcane endophytic and plant-growth promoting traits of G. diazotrophicus. An accessory component of at least 851 genes distributed in genome islands was identified, and was most likely acquired by horizontal gene transfer. This portion of the genome has likely contributed to adaptation to the plant habitat.ConclusionThe genome data offer an important resource of information that can be used to manipulate plant/bacterium interactions with the aim of improving sugarcane crop production and other biotechnological applications.
With the increasing number of pets in home the human-animal relationship is increasingly close and care about control disease growing. Ivermectin (IVM) is frequently used because its proven safety. IVM is recommended for the treatment of demodectic scabies and prevention of heartworm in dogs, but informally is extremely used to control of Ctenocephalides felis felis and Rhipicephalus sanguineus. The aim of this study is evaluate the use of IVM in dogs, by the oral route at 0.6μg/kg dose, against experimental infection of these parasites using the construction of the plasma concentration curve and efficacy study. A IVM quantification method in canine plasma using HPLC-FL was developed and validated based on RE n°899/03 ANVISA. The samples collected during the efficacy test was analyzed by this validated method and prove C of 350ng/mL at 4h (t) and AUC of 8411ng/h/mL. Spite of formulation have shown good absorption, the highest efficiency values found for Rhipcephalus sanguineus and Ctenocephalides felis felis were very low, 35% and 67% respectively, demonstrating this not be the most appropriate treatment for the control of these parasites.
Fipronil is a phenylpyrazole class insecticide. It is widely used as an insecticide in agriculture and in the control of ectoparasites in veterinary medicine. The application of fipronil in an injectable form (subcutaneously) becomes an innovation, since there is no commercially available preparation containing fipronil herein. The present study aimed at fipronil usage, applied subcutaneously in cattle, to control Rhipicephalus microplus. The assessing criteria used in the research have been the construction of the plasma concentration curve and efficacy studies. A method using High Performance Liquid Chromatograph with ultraviolet detection was developed for determination of fipronil in bovine plasma samples, providing a fast and simple process with good reproducibility and low limit of quantification. The validation of the analytical method showed linearity, selectivity, precision, accuracy, sensitivity and stability, thus proving it as suitable for routine analysis. This method showed to be an important investigative tool in the analysis of fipronil plasma concentration in cattle. Fipronil administered via subcutaneous in bovine reached the systemic circulation (Cmax=378.06±137.44 ng/mL), was quickly absorbed (t(max)=10±0.87 h), and its elimination occurred slowly (t(1/2)=12 days), while maintaining quantifiable blood plasma levels (23.79±12.16 ng/mL) for up to 21 days after the treatment with a 1 mg/kg dosage. The in vivo efficacy tests proved that fipronil applied subcutaneously in a single dose of 1 mg/kg in cattle exhibited a mean efficacy of 82.41% against R. microplus. The potential of subcutaneous injection as an alternative treatment route in cattle encourage the development of an injectable formulation of fipronil.
Increased human-pet interactions have led to concerns related to the prevention and treatment of ectoparasite infestations. Fipronil (FIP) is a widely used ectoparasiticide in veterinary medicine available for topical administration; however, its use may cause damage to the owners and the environment. The aim of the study was to develop immediate-release tablets of FIP, as well as to determine its pharmacokinetic properties after oral administration in beagle dogs. The prepared FIP tablets were evaluated for precompression (angle of repose, speed flow, and Carr's index) and post-compression (weight variation, friability, thickness, hardness, disintegration time, and dissolution rate) parameters. Orally administered FIP at a dose of 2 mg/kg was rapidly absorbed with C máx of 3.13 ± 1.39 μg/mL at 1.83 ± 0.40 h post treatment (P.T.) and metabolized with 1.27 ± 1.04 μg/mL at 2.33 ± 0.82 h P.T. for fipronil sulfone (SULF) (the primary metabolite). The elimination of FIP and SULF occurred slowly and had maintained quantifiable plasma levels in the blood for up to 28 days P.T. The goal of the study is aligned with the concept of One Health, which aims to collaboratively achieve the best health for people, animals, and the environment. Therefore, the use of FIP tablets for the control of ectoparasites in dogs may be a safer alternative for owners and the environment.
Recebido em 7/3/12; aceito em 9/7/12; publicado na web em 25/9/12 A fast and efficient method has been developed and validated for the determination of fipronil in bovine plasma. Samples were subjected to solid-phase extraction (SPE) followed by reversed phase liquid chromatography (LC) separation, using acetonitrile/ water (60:40 v/v) as the mobile phase with a flow rate of 1.0 mL/min and ultraviolet (UV) detection at 210 nm. Ethiprole was used as the internal standard (IS). The method was found to be linear over the range 5-500 ng/mL (r = 0.999). The limit of quantitation (LOQ) was validated at 5 ng/mL. The method was successfully applied to monitor plasma concentrations following subcutaneous administration of fipronil in cattle.
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