A comparison of toxicant-induced succession for five antifouling compounds on marine periphyton in SWIFT microcosms

Five antifouling biocides, chlorothalonile, dichlofluanide, medetomidine, tolylfluanide, and zinc pyrithione, were evaluated regarding their effect on the composition of the periphyton community and the subsequent toxicant-induced succession (TIS). The periphyton communities were exposed in a semi-static setting for 96 h using a SWIFT microcosm. As a measure of community composition, pigment profiles from the exposed communities were used as effect indicators and compared with unexposed parts of the same community using the Bray–Curtis dissimilarity index. Chlorothalonile caused changes in the community starting at 85 μg l^?1 while dichlofluanide had no effect even at the highest concentrations used, 810 μg l^?1. The related substance tolylfluanide only affected the community composition at 2700 μg l^?1. Medetomidine had a different response curve with a small effect on the community composition at 0.8 μg l^?1 which then disappeared only to reappear at 240 μg l^?1. Zinc pyrithione had the largest effect on the periphyton community with changes starting at 10 μg l^?1 and no detectable pigments at 100 μg l^?1. The changes in the community composition for the five substances were also compared using multidimensional scaling. When all substances were analyzed and plotted together, chlorothalonile, dichlofluanide, medetomidine, and tolylfluanide showed surprisingly similar effects compared to zinc pyrithione that gave very different TIS. However, when only chlorothalonile, dichlofluanide, and tolylfluanide were plotted together, clear differences in TIS between the three toxicants were revealed. Dichlofluanide only induced small effects, while concentration-dependent TIS trajectories for chlorothalonile and tolylfluanide took off in opposite directions indicating very different responses of the periphyton communities. This study demonstrates that substances with a similar chemical structure and mechanisms of action can have different effects on the community composition. With the exception of zinc pyrithione, none of the recorded effect levels were at concentrations reported from marine environments so far.     

Spontaneous Super-Swarming Derivatives of Azospirillum brasilense Sp245 have Different DNA Profiles and Behavior in the Presence of Various Nitrogen Sources

Azospirillum brasilense swims in liquid environments and swarms in semisolid media. Five variants of A. brasilense Sp245, Sp245.P1–Sp245.P5, which swarmed faster than Sp245 in a semisolid malate–salt medium, have been isolated. In Sp245.P1–Sp245.P4, a new megaplasmid was revealed instead of an indigenous 85-MDa plasmid (p85). By polymerase chain reactions (PCR) with primers to the segments of p85 important for proper bacterial motility/flagellation and for dissimilatory nitrite and NO reduction, that DNA of p85 was found retained by all the variants. In ERIC- and RAPD-PCR, microdiversity between the total DNAs of Sp245 and its variants was detected. Interstrain differences in growth characteristics in liquid peptone–succinate–salt medium with KNO₃ or KNO₂ and in KNO₂ production/consumption were revealed. Although all the variants swam and swarmed faster than Sp245 in the medium supplemented with NH₄Cl or KNO₃, not all of them could do so in MPSS with KNO₂.     

A calibration method to generate seasonal hatching profiles for the fiddler crabs Uca pugnax (Smith, 1870) and Uca minax (LeConte, 1855) (Decapoda : Brachyura)

A statistical method for generating seasonal egg hatching profiles is applied to the brachyuran crabs Uca pugnax (Smith) and U. minax (LeConte) in New Jersey. Calibration experiments quantified the time course of egg development, using serial egg sampling at 1- to 2- day intervals from marked ovigerous females maintained in cages in the field. Egg stage was ranked from 1 to 10, based on morphological changes during development. Equations for predicting the number of days remaining until hatching from egg developmental stage were obtained from the calibration experiments, using stepwise polynomial regression. To cover the reproductive season, three consecutive calibration experiments using 15 or more females were run for U. pugnax; two for U. minax. Significant seasonal differences in the time course of egg development were detected. Weekly collections of females for each species were made; the date of larval release for each ovigerous female was predicted from the proximate calibration equation, yielding weekly hatching profiles. Weekly hatching profiles were summed to obtain seasonal hatching profiles. The average number of broods produced per female over the reproductive season was 1.9 for U. pugnax and 1.6 for U. minax. Hatching peaks for both species were associated with spring tides. The merits of this and other methods used to estimate daily variation in egg hatching of crabs are discussed.     

Biosystematics of alkaliphilic streptomycetes isolated from seven locations across a beach and dune sand system

Alkaliphilic streptomycetes were isolated from composite sand samples collected from six out of seven locations across a beach and dune sand system using starch-casein-nitrate agar supplemented with cycloheximide and buffered to pH 10.5. The isolates had colonial and chemotaxonomic properties consistent with their classification in the genus Streptomyces. They were assigned to 49 multimembered and 114 single-membered colour-groups given their ability to produce pigments on oatmeal and peptone-yeast-extract-iron agars and to corresponding taxa based on whole-genome rep-PCR banding patterns. Twenty-four isolates representing the colour and rep-PCR groups grew well from pH 5 to 11, and optimally at pH 9, as did phylogenetically close members of the Streptomyces griseus 16S rRNA gene clade. One hundred and twelve representative alkaliphilic streptomycetes formed a heterogeneous but distinct clade in the Streptomyces 16S rRNA gene tree. A 3-dimensional representation of 16S rRNA sequence data showed that the alkaliphilic streptomycetes formed a distinct group in multidimensional taxospace. It is evident that alkaliphilic streptomycetes are common in the beach and dune sand system and that representatives of this community form new centers of taxonomic variation within the genus Streptomyces that can be equated with species. GenBank accession numbers for the 16S rRNA gene sequences for the strains of the alkaliphilic streptomycetes Bd 095, Bd 064, Bd 077, Bd 013, Bd 108, Bd 088, Bd 012, Bd 187, Bd 128, Bd 174, Bd 167, Lt 005, Lt 006, Fd 015, Bd 099, Bd 059, Bd 159, Ht 015, Md 005, Ht 020, Bd 205, Md 063, Fd 004, Md 039 and Bd 092 are EU477215, EU477216, EU477217, EU477218, EU477219, EU477220, EU477221, EU477222, EU477223, EU477224, EU477225, EU477226, EU477227, EU477228, EU477229, EU477230, EU477231, EU477232, EU477233, EU477234, EU477235, EU477236, EU477237, EU477238 and EU477257, respectively.     

Distribution and Risk Assessment of Heavy Metals in Sediments of the Pearl River Estuary,Southern China

Sediment cores collected at six freshwater discharge outlets of the Pearl River Estuary were analyzed for metal (Al, Fe, Cr, Pb, and Cd) concentration and risk assessment. The contents of Cr, Pb, Fe, and Al in these samples were measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES), and Cd was measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The result showed that the concentration ranged (in mg/kg dry weight) as follows: Al, 7,880–97,700; Fe, 30,100–51,900; Cd, 0.23–1.09; Cr, 48.7–109; Pb, 22.3–70.2; respectively. The sediments of different particle size were measured and the result showed that the pelitic fraction (< 0.05 mm) was more enriched in heavy metals. Cd and Pb showed a similar spatial distribution in the sediment, which reflected similar anthropogenic origins. In addition, the temporal distribution of Cr, Pb, and Cd showed a light increase from the 1970s to the 2000s. Furthermore, the enrichment factor (EF) and the geo-accumulation index (I_geo) were used to comprehensively evaluate the pollution degree of heavy metals. The findings will be useful in proposing measures for strategic environmental control in estuaries.     

Architecture and assembly of a divergent member of the ParM family of bacterial actin-like proteins

Eubacteria and archaea contain a variety of actin-like proteins (ALPs) that form filaments with surprisingly diverse architectures, assembly dynamics, and cellular functions. Although there is much data supporting differences between ALP families, there is little data regarding conservation of structure and function within these families. We asked whether the filament architecture and biochemical properties of the best-understood prokaryotic actin, ParM from plasmid R1, are conserved in a divergent member of the ParM family from plasmid pB171. Previous work demonstrated that R1 ParM assembles into filaments that are structurally distinct from actin and the other characterized ALPs. They also display three biophysical properties thought to be essential for DNA segregation: 1) rapid spontaneous nucleation, 2) symmetrical elongation, and 3) dynamic instability. We used microscopic and biophysical techniques to compare and contrast the architecture and assembly of these related proteins. Despite being only 41% identical, R1 and pB171 ParMs polymerize into nearly identical filaments with similar assembly dynamics. Conservation of the core assembly properties argues for their importance in ParM-mediated DNA segregation and suggests that divergent DNA-segregating ALPs with different assembly properties operate via different mechanisms.     

PsiB polypeptide prevents activation of RecA protein in Escherichia coli

Summary We further characterize a novel plasmid function preventing SOS induction called Psi (Plasmid SOS Inhibition). We show that Psi function is expressed by psiB, a gene located at coordinate 54.9 of plasmid R6-5 and near oriT, the origin of conjugal transfer. Deletions and amber mutations of the psiB gene permitted us to demonstrate that PsiB polypeptide (apparent molecular weight, 12 kDa) is responsible for Psi function. PsiB protein prevents recA730-promoted mutagenesis and intra-chromosomal recombination but not recombination following conjugation. Overproduction of PsiB protein sensitizes the host cell to UV irradiation. We propose that PsiB polypeptide has an anti-SOS action by inhibiting activation of RecA protein, thus preventing the occurrence of LexA-controlled functions.     

Efficient genetic approaches for improvement of plasmid based expression of recombinant protein in Escherichia coli: A review

During the past two decades, there has been an explosion of new knowledge and techniques in the field of recombinant protein expression. However, over-expression of “difficult to express proteins” with therapeutic importance continues to be a challenging task for successful commercialization of these proteins. With the emergence of the bio-similar market, enhancing the efficiencies of the production process has become a critical factor in the commercial viability of novel products. Despite the availability of numerous technological advancements, recombinant protein expression in Escherichia coli remains difficult. Therefore, addressing upstream bottlenecks in combination with genetically modified expression hosts could be a viable strategy to enhance production. Problems like poor expression, plasmid instability, protein aggregation, protein degradation, and metabolic stress associated with recombinant protein production need special consideration during bioprocess development at bioreactor level. However, a comprehensive universal strategy for attaining efficient expression in E. coli seems unrealistic and must be resolved empirically. In this review, we have discussed some common problems and their apparent solutions for plasmids based recombinant gene expression in E. coli.