Pseudoalteromonas is widespread in various marine environments, and most strains can affect invertebrate larval settlement and metamorphosis by forming biofilms. However, the impact and the molecular basis of population diversification occurring in Pseudoalteromonas biofilms are poorly understood. Here, we show that morphological diversification is prevalent in Pseudoalteromonas species during biofilm formation. Two types of genetic variants, wrinkled (frequency of 12 ± 5 %) and translucent (frequency of 5 ± 3 %), were found in Pseudoalteromonas lipolytica biofilms. The inducing activities of biofilms formed by the two variants on larval settlement and metamorphosis of the mussel Mytilus coruscus were significantly decreased, suggesting strong antifouling activities. Using whole-genome re-sequencing combined with genetic manipulation, two genes were identified to be responsible for the morphology alternations. A nonsense mutation in AT00_08765 led to a wrinkled morphology due to the overproduction of cellulose, whereas a point mutation in AT00_17125 led to a translucent morphology via a reduction in capsular polysaccharide production. Taken together, the results suggest that the microbial behavior on larval settlement and metamorphosis in marine environment could be affected by the self-generated variants generated during the formation of marine biofilms, thereby rendering potential application in biocontrol of marine biofouling.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-015-6865-x) contains supplementary material, which is available to authorized users.

Background The hard-shelled mussel (Mytilus coruscus) is widely distributed in the temperate seas of East Asia and is an important commercial bivalve in China. Chromosome-level genome information of this species will contribute not only to the development of hard-shelled mussel genetic breeding but also to studies on larval ecology, climate change biology, marine biology, aquaculture, biofouling, and antifouling. Findings We applied a combination of Illumina sequencing, Oxford Nanopore Technologies sequencing, and high-throughput chromosome conformation capture technologies to construct a chromosome-level genome of the hard-shelled mussel, with a total length of 1.57 Gb and a median contig length of 1.49 Mb. Approximately 90.9% of the assemblies were anchored to 14 linkage groups. We assayed the genome completeness using BUSCO. In the metazoan dataset, the present assemblies have 89.4% complete, 1.9% incomplete, and 8.7% missing BUSCOs. Gene modeling enabled the annotation of 37,478 protein-coding genes and 26,917 non-coding RNA loci. Phylogenetic analysis showed that M. coruscus is the sister taxon to the clade including Modiolus philippinarum and Bathymodiolus platifrons. Conserved chromosome synteny was observed between hard-shelled mussel and king scallop, suggesting that this is shared ancestrally. Transcriptomic profiling indicated that the pathways of catecholamine biosynthesis and adrenergic signaling in cardiomyocytes might be involved in metamorphosis. Conclusions The chromosome-level assembly of the hard-shelled mussel genome will provide novel insights into mussel genome evolution and serve as a fundamental platform for studies regarding the planktonic-sessile transition, genetic diversity, and genomic breeding of this bivalve.

Molecular cloning and nucleotide sequencing of cDNA encoding Bombyx mori nitric oxide synthase (BmNOS) was conducted to analyse its possible role in insect immunity. The amino acid sequence deduced from the BmNOS cDNA showed 84%, 54% and 53% identity with those of NOSs from Manduca sexta, Drosophila melanogaster and Rhodonius prolixus. Recombinant BmNOS produced in insect cells using baculovirus was found to require NADPH, Ca2+, calmodulin and tetrahydrobiopterin (BH4) for its activity. The BmNOS gene was constitutively expressed at a low level in the larval fat body, haemocyte, Malpighian tubule and midgut, and adult antenna, and induced strongly in the fat body by lipopolysaccharide (LPS), suggesting that the BmNOS gene plays different physiological roles in different tissues. Injection of NO donors that produce NO in vivo induced the gene expression of an antibacterial peptide, cecropin B, strongly suggesting that NO produced by BmNOS following LPS stimulation is involved in signal transduction as a signalling molecule for immune gene expression.

ABSTRACT. This study aimed to provide additional anatomical information for axillary lymph node dissection (ALND) through in vivo anatomy studies of intercostobrachial nerve (ICBN) preservation in order to provide theoretical and practical experience for clinicians. A total of 156 patients with breast cancer underwent ALND at the Department of Gynecology of Baotou Tumor Hospital between June 2009 and March 2010. The origin, destination, main source, length, branch type, and direction of ICBN in axilla were observed, as well as its relationship with adjacent major blood vessels and nerves within the axilla. There were 120 cases of single trunk, 23 cases of double trunks, 9 cases of multiple trunks, and 4 cases without trunks in 156 patients with ICBN preservation. The transverse diameter at the origin of the ICBN was 1.89 ± 0.44 mm with a length of 94.45 ± 12.08 mm; the distances were 77.19 ± 21.04 mm, 29.34 ± 6.73 mm, 90.04 ± 13.13 mm, and 28.63 ± 13.01 mm from origin to the inferior margin at the midpoint of the clavicle, inferior margin of the axillary vein, the bottom of axilla, and branch point, respectively. The identification, dissection, and preservation of ICBN was simple and easy in a modified radical mastectomy for breast cancer and breast-conserving surgery, which only took 10-20 min, but effectively reduced the incidence of post-mastectomy pain syndrome and significantly improved the quality of life for patients after surgery.

This study investigated the effect of carbon nanotubes (CNTs) and titanium dioxide (TiO2) incorporated in PDMS on biofilm formation and plantigrade settlement of Mytilus coruscus. TiO2 increased bacterial density, and CNTs also increased bacterial density but reduced diatom density in biofilms after 28 days. Further analysis was conducted between bacterial communities on glass, PDMS, CNTs (0.5 wt%) and TiO2 (7.5 wt%). ANOSIM analysis revealed significant differences (R > 0.9) between seven, 14, 21 and 28 day-old bacterial communities. MiSeq sequencing showed that CNTs and TiO2 impacted the composition of 28 day-old bacterial communities by increasing the abundance of Proteobacteria and decreasing the abundance of Bacteroidetes. The maximum decreased settlement rate in 28 day-old biofilms on CNTs and TiO2 was > 50% in comparison to those on glass and PDMS. Thus, CNTs and TiO2 incorporated in PDMS altered the biomass and community composition of biofilms, and subsequently decreased mussel settlement.

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