The sps Genes Encode an Original Legionaminic Acid Pathway Required for Crust Assembly in Bacillus subtilis.

The crust is the outermost spore layer of most Bacillus strains devoid of an exosporium. This outermost layer, composed of both proteins and carbohydrates, plays a major role in the adhesion and spreading of spores into the environment. Recent studies have identified several crust proteins and have provided insights about their organization at the spore surface. However, although carbohydrates are known to participate in adhesion, little is known about their composition, structure, and localization. In this study, we showed that the spore surface of Bacillus subtilis is covered with legionaminic acid (Leg), a nine-carbon backbone nonulosonic acid known to decorate the flagellin of the human pathogens Helicobacter pylori and Campylobacter jejuni We demonstrated that the spsC, spsD, spsE, spsG, and spsM genes of Bacillus subtilis are required for Leg biosynthesis during sporulation, while the spsF gene is required for Leg transfer from the mother cell to the surface of the forespore. We also characterized the activity of SpsM and highlighted an original Leg biosynthesis pathway in B. subtilis Finally, we demonstrated that Leg is required for the assembly of the crust around the spores, and we showed that in the absence of Leg, spores were more adherent to stainless steel probably because of their reduced hydrophilicity and charge.IMPORTANCEBacillus species are a major economic and food safety concern of the food industry because of their food spoilage-causing capability and persistence. Their persistence is mainly due to their ability to form highly resistant spores adhering to the surfaces of industrial equipment. Spores of the Bacillus subtilis group are surrounded by the crust, a superficial layer which plays a key role in their adhesion properties. However, knowledge of the composition and structure of this layer remains incomplete. Here, for the first time, we identified a nonulosonic acid (Leg) at the surfaces of bacterial spores (B. subtilis). We uncovered a novel Leg biosynthesis pathway, and we demonstrated that Leg is required for proper crust assembly. This work contributes to the description of the structure and composition of Bacillus spores which has been under way for decades, and it provides keys to understanding the importance of carbohydrates in Bacillus adhesion and persistence in the food industry.

Short-term exposure to gold nanoparticle suspension impairs swimming behavior in a widespread calanoid copepod.

Calanoid copepods play an important role in the functioning of marine and brackish ecosystems. Information is scarce on the behavioral toxicity of engineered nanoparticles to these abundant planktonic organisms. We assessed the effects of short-term exposure to nonfunctionalized gold nanoparticles on the swimming behavior of the widespread estuarine copepod Eurytemora affinis. By means of three-dimensional particle tracking velocimetry, we reconstructed the trajectories of males, ovigerous and non-ovigerous females. We quantified changes in their swimming activity and in the kinematics and geometrical properties of their motion, three important descriptors of the motility patterns of zooplankters. In females, exposure to gold nanoparticles in suspension (11.4 µg L-1) for 30 min caused depressed activity and lower velocity and acceleration, whereas the same exposure caused minimal effects in males. This response differs clearly from the hyperactive behavior that is commonly observed in zooplankters exposed to pollutants, and from the generally lower sensitivity of female copepods to toxicants. Accumulation of gold nanoparticles on the external appendages was not observed, precluding mechanical effects. Only very few nanoparticles appeared sporadically in the inner part of the gut in some samples, either as aggregates or as isolated nanoparticles, which does not suggest systemic toxicity resulting from pronounced ingestion. Hence, the precise mechanisms underlying the behavioral toxicity observed here remain to be elucidated. These results demonstrate that gold nanoparticles can induce marked behavioral alterations at very low concentration and short exposure duration. They illustrate the applicability of swimming behavior as a suitable and sensitive endpoint for investigating the toxicity of nanomaterials present in estuarine and marine environments. Changes in swimming behavior may impair the ability of planktonic copepods to interact with their environment and with other organisms, with possible impacts on population dynamics and community structure.

Cumulative effect of zinc oxide and titanium oxide nanoparticles on growth and chlorophyll a content of Picochlorum sp.

The use of nanoparticles (NPs) is of increasing significance due to their large potential for various applications. Great attention should be paid on the possible impacts of nanoparticles on the environment as large amounts of them may reach the environment by accident or voluntarily. Marine algae are potential organisms for usage in nanopollution bioremediation in aquatic system, because of their ability to adapt to long exposure to NPs. Thus, it is of prime importance to study the possible interactions of different NPs with microalgae in assessing their potential environmental risks. Most studies on potential environmental effects of ZnO and TiO2 NPs have been performed independently and following the widely accepted, standardized test systems, which had been developed for the characterization of chemicals. In this study, we have examined the cumulative effect of ZnO and TiO2 NPs on Picochlorum sp. in addition to the individual effects of these NPs over 32 days. Our results indicate that the toxicity and availability of NPs to marine algae are reduced by their aggregation and sedimentation. NPs are found to have a negative effect on algal growth and chlorophyll a concentration during the early growth stages. In contrast, the case is reversed during the late growth stages. There is no significant difference between the effect of the NPs when they are used separately and when both ZnO and TiO2 are used together in the test (P > 0.05).

Effect of magnetic iron oxide (Fe3O4) nanoparticles on the growth and photosynthetic pigment content of Picochlorum sp.

Magnetite iron oxide (Fe3O4) nanoparticles (NPs) are key materials applied in many different fields of modern technology. The potential environmental impact of these NPs is of great concern. In this study, initially the effect of Fe3O4 NPs size (20 and 40 nm) as well as bulk (>100 nm) at 200 mg L(-1) on Picochlorum sp. (Trebouxiophyceae, Chlorophyta) is investigated during the different growth phases. The most inhibitory NPs were then chosen to assess their effects at different concentrations. The 20 nm NPs at 200 mg L(-1) were found to significantly reduce the viable cell concentration and chlorophyll a content during the exponential growth phase compared to the other particle sizes. However, the 20 nm NPs at different concentrations were found to promote algal growth during the late growth stages (stationary and decline phases) compared to the control. Additionally, algae were found to accelerate the aggregation and sedimentation of nanoparticles into the medium and therefore can be considered as potential organisms for bioremediation of nano-pollution.