RESUMEN
Exiguobacterium antarcticum strain B7 is a psychrophilic Gram-positive bacterium that possesses enzymes that can be used for several biotechnological applications. However, many proteins from its genome are considered hypothetical proteins (HPs). These functionally unknown proteins may indicate important functions regarding the biological role of this bacterium, and the use of bioinformatics tools can assist in the biological understanding of this organism through functional annotation analysis. Thus, our study aimed to assign functions to proteins previously described as HPs, present in the genome of E. antarcticum B7. We used an extensive in silico workflow combining several bioinformatics tools for function annotation, sub-cellular localization and physicochemical characterization, three-dimensional structure determination, and protein-protein interactions. This genome contains 2772 genes, of which 765 CDS were annotated as HPs. The amino acid sequences of all HPs were submitted to our workflow and we successfully attributed function to 132 HPs. We identified 11 proteins that play important roles in the mechanisms of adaptation to adverse environments, such as flagellar biosynthesis, biofilm formation, carotenoids biosynthesis, and others. In addition, three predicted HPs are possibly related to arsenic tolerance. Through an in vitro assay, we verified that E. antarcticum B7 can grow at high concentrations of this metal. The approach used was important to precisely assign function to proteins from diverse classes and to infer relationships with proteins with functions already described in the literature. This approach aims to produce a better understanding of the mechanism by which this bacterium adapts to extreme environments and to the finding of targets with biotechnological interest.
Asunto(s)
Adaptación Fisiológica , Arsénico/toxicidad , Bacillaceae/fisiología , Proteínas Bacterianas/fisiología , Anotación de Secuencia Molecular , Proteínas Bacterianas/genética , Biotecnología/métodos , Biología Computacional , Ambientes Extremos , Genes Bacterianos/genética , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: The control of Aedes aegypti is usually based on chemical insecticides, but the overuse of these compounds has led to increased resistance. The binary toxin produced by Lysinibacillus sphaericus in the final stages of sporulation is used for mosquito control due to its specificity against the culicid larvae; however, it has been proved that Ae. aegypti is refractory for this toxin. Currently, there is no evidence of the use of L. sphaericus vegetative cells for mosquito biocontrol. Therefore, in this study, the vegetative cells of three L. sphaericus strains were assessed against a field-collected Ae. aegypti, resistant to temephos, and the reference Rockefeller strain. RESULTS: Vegetative cells of L. sphaericus 2362, III(3)7 and OT4b.25 produced between 90% and 100% of larvae mortality in the reference Rockefeller strain. Effective concentrations of each L. sphaericus strain for the four larval stages ranged from 1.4 to 2 × 107 CFU/ml. Likewise, a consortium of L. sphaericus assessed against a field-collected Ae. aegypti resistant to temephos and the Rockefeller strain caused 90% of larvae mortality. Concentrations of L. sphaericus consortium that resulted in larvae mortality of field-collected and Rockefeller Ae. aegypti ranged from 1.7 to 2.5 × 107 CFU/ml. The vegetative cells of L. sphaericus have no effect on the Ae. aegypti eggs and pupae. CONCLUSIONS: The vegetative cells of L. sphaericus are effective against Ae. aegypti larvae, meaning that it could be used in the biological control of these mosquito species. Since the L. sphaericus consortium was effective against temephos-resistant Ae. aegypti, vegetative cells could be an alternative to overcome insecticide-resistant populations. Further studies, should be conducted to reveal the mode of action and the toxic principle of L. sphaericus vegetative cells.
Asunto(s)
Aedes/microbiología , Bacillaceae/fisiología , Agentes de Control Biológico , Larva/microbiología , Control de Mosquitos , Animales , Bacillaceae/química , Resistencia a los Insecticidas , Larva/efectos de los fármacos , TemefósRESUMEN
OBJECTIVE: Assessment of the bacterium L. sphaericus as a dual-action candidate for biological control of mosquito-borne diseases and bioremediation of toxic metals. METHODS: Larvae of the mosquito, C. quinquefasciatus, were first evaluated for metal tolerance and then exposed to 5 ppm cadmium, chromium, arsenic, and lead in assays together with seven strains of L. sphaericus. A probit regression analysis was used to estimate the LC(50) of Cd, Cr, As, and Pb to C. quinquefasciatus. An analysis of covariance and multifactorial ANOVA examined the metal biosorption and larvicidal properties of the seven strains of L. sphaericus. RESULTS: We found that L. sphaericus adsorbed the toxic metal ions and was toxic against mosquito larvae. The L. sphaericus strain III(3)7 resulted in a larvae mortality of over 80% for all the tested metals. This strain also exhibited the capacity to adsorb 76% of arsenic, 32% of lead, 25% of chromium, and 7% of cadmium. CONCLUSION: This study found combined metal adsorption and larval toxicity associated with three strains of L. sphaericus [III(3)7, OT4b.31, and CBAM5]. This suggests that a combination of these strains shows strong dual potential for biological control of mosquitos in heavy metal-contaminated areas and remediate the heavy metal contamination as well.
Asunto(s)
Bacillaceae/fisiología , Culicidae/microbiología , Metales Pesados/metabolismo , Contaminantes Químicos del Agua/metabolismo , Animales , Interacciones Huésped-Patógeno , Insectos Vectores , Larva/microbiología , Metales Pesados/toxicidad , Contaminantes Químicos del Agua/toxicidadRESUMEN
Lysinibacillus sphaericus is a spore-forming bacterium used in the biological control of mosquitoes and in bioremediation. Mosquito larvae exposed to heavy metals are tolerant to concentrations above the permissible limit for industrial residual waters. In this work, we characterize 51 L. sphaericus strains for metal tolerance and larvicidal activity against Culex quinquefasciatus. Lysinibacillus sphaericus OT4b.2, OT4b.20, OT4b.25, OT4b.26 and OT4b.58 were as toxic as the spores of the reference strain 2362 against C. quinquefasciatus larvae. 19 Mosquito-pathogenic L. sphaericus strains and 6 non-pathogenic strains were able to grow in arsenate, hexavalent chromium and/or lead. 16S rRNA gene sequences and phylogenetic analyses clustered 84 % of the metal-tolerant strains in L. sphaericus group 1, which encompasses the mosquitocidal strains. The larvicidal activity of vegetative and sporulated cells and its high tolerance to arsenate, hexavalent chromium and lead indicate that L. sphaericus OT4b.26 is a strong candidate for further studies examining its potential for biological control of mosquitoes in waters contaminated with metals.
Asunto(s)
Arseniatos/metabolismo , Bacillaceae/fisiología , Cromo/metabolismo , Culex/crecimiento & desarrollo , Culex/microbiología , Plomo/metabolismo , Control Biológico de Vectores , Animales , Bacillaceae/clasificación , Bacillaceae/genética , Bacillaceae/aislamiento & purificación , Escarabajos/microbiología , Larva/crecimiento & desarrollo , Larva/microbiología , Datos de Secuencia Molecular , Filogenia , Microbiología del SueloRESUMEN
The evolutionary history and ecological differentiation of the genus Exiguobacterium was characterized within natural communities from the Cuatro Cienegas Basin, Mexico. Exiguobacterium comprises both halophilic and alkaliphilic bacteria that are abundant among the aquatic systems of the Cuatro Cienegas Basin. We obtained complete sequences of the 16srRNA gene and partial sequences of four housekeeping genes (citC, rpoB, recA and hsp70) in 183 Exiguobacterium isolates retrieved from distinct aquatic systems. We defined three main phylogroups that are closely related to marine and thermophilic species of the genus. These phylogroups were neither specific to a given aquatic system nor to a particular salinity. Phylogenetic reconstruction indicated the presence of several small clusters within the phylogroups. These clusters consisted of isolates predominantly retrieved from sediment or water. Unifrac and AdaptML analyses confirmed this observation, pointing towards a clear pattern of differentiation linked to either sediment or water habitats. Our results are in line with the concept that niche differentiation is one of the main factors shaping prokaryotic populations and leading to evolutionary divergence.
Asunto(s)
Bacillaceae/clasificación , Bacillaceae/fisiología , Ecosistema , Filogenia , Microbiología del Suelo , Microbiología del Agua , Bacillaceae/genética , Bacillaceae/metabolismo , Biodiversidad , Ecología , México , SalinidadRESUMEN
Tellurite exerts a deleterious effect on a number of small molecules containing sulfur moieties that have a recognized role in cellular oxidative stress. Because cysteine is involved in the biosynthesis of glutathione and other sulfur-containing compounds, we investigated the expression of Geobacillus stearothermophilus V cysteine-related genes cobA, cysK, and iscS and Escherichia coli cysteine regulon genes under conditions that included the addition of K2TeO3 to the culture medium. Results showed that cell tolerance to tellurite correlates with the expression level of the cysteine metabolic genes and that these genes are up-regulated when tellurite is present in the growth medium.
Asunto(s)
Antibacterianos/farmacología , Bacillaceae/efectos de los fármacos , Cisteína/genética , Escherichia coli/efectos de los fármacos , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Telurio/farmacología , Transferasas Alquil y Aril/biosíntesis , Bacillaceae/genética , Bacillaceae/fisiología , Proteínas Bacterianas/biosíntesis , Liasas de Carbono-Azufre/biosíntesis , Cisteína/metabolismo , Cisteína Sintasa/biosíntesis , Escherichia coli/genética , Escherichia coli/fisiología , Proteínas de Escherichia coli/biosíntesis , RegulónRESUMEN
Spore suspensions of a pure culture of Alicyclobacillus acidoterrestris DSM 2498 were submitted to different heat treatments (60 degrees C for 60 min, 60 degrees C for 30 min, 70 degrees C for 20 min, 80 degrees C for 5 min, 80 degrees C for 10 min, 80 degrees C for 30 min, and boiling for 5 min) to determine the best activation conditions in orange juice. The best treatment for spore activation was shown to be 70 degrees C/20 min. Seventy-five samples of concentrated orange juice from 11 different suppliers were examined for the presence of thermophilic acid-tolerant spore formers by the most probable number technique using Bacillus acidocaldarius medium (BAM broth) and incubation at 44 degrees C for 5 days after a prior spore activation. After incubation, isolation was carried out using BAM agar medium incubating at 44 degrees C for 5 days. Typical colonies were submitted to a microscopic examination, evaluation for the presence of spores, and various biochemical tests. Of the orange juice samples examined, 14.7% were found to be positive for Alicyclobacillus. The thermal death time open tube method was used to determine the heat resistance of the spores of strains confirmed as being Alicyclobacillus. The D-values determined were in the range from 60.8 to 94.5 min at 85 degrees C, 10.0 to 20.6 min at 90 degrees C, and 2.5 to 8.7 min at 95 degrees C. The z-values were between 7.2 degrees C and 11.3 degrees C. The results demonstrated the occurrence of Alicyclobacillus in orange juice and the high heat resistance of the spores that could survive the heat treatments normally applied in the processing of orange juice.