Characterization of a Novel Conus bandanus Conopeptide Belonging to the M-Superfamily Containing Bromotryptophan

A novel conotoxin (conopeptide) was biochemically characterized from the crude venom of the molluscivorous marine snail, Conus bandanus (Hwass in Bruguière, 1792), collected in the south-central coast of Vietnam. The peptide was identified by screening bromotryptophan from chromatographic fractions of the crude venom. Tandem mass spectrometry techniques were used to detect and localize different post-translational modifications (PTMs) present in the BnIIID conopeptide. The sequence was confirmed by Edman’s degradation and mass spectrometry revealing that the purified BnIIID conopeptide had 15 amino acid residues, with six cysteines at positions 1, 2, 7, 11, 13, and 14, and three PTMs: bromotryptophan, γ-carboxy glutamate, and amidated aspartic acid, at positions “4”, “5”, and “15”, respectively. The BnIIID peptide was synthesized for comparison with the native peptide. Homology comparison with conopeptides having the III-cysteine framework (–CCx₁x₂x₃x₄Cx₁x₂x₃Cx₁CC–) revealed that BnIIID belongs to the M-1 family of conotoxins. This is the first report of a member of the M-superfamily containing bromotryptophan as PTM.     

Multiple Beneficial Lipids Including Lecithin Detected in the Edible Invasive Mollusk Crepidula fornicata from the French Northeastern Atlantic Coast

The invasive mollusk Crepidula fornicata, occurring in large amounts in bays along the French Northeastern Atlantic coasts, may have huge environmental effects in highly productive ecosystems where shellfish are exploited. The present study aims at determining the potential economic value of this marine species in terms of exploitable substances with high added value. Lipid content and phospholipid (PL) composition of this mollusk collected on the Bourgneuf Bay were studied through four seasons. Winter specimens contained the highest lipid levels (5.3% dry weight), including 69% of PLs. Phosphatidylcholine (PC) was the major PL class all year, accounting for 63.9% to 88.9% of total PLs. Consequently, the winter specimens were then investigated for PL fatty acids (FAs), and free sterols. Dimethylacetals (DMAs) were present (10.7% of PL FA + DMA mixture) revealing the occurrence of plasmalogens. More than forty FAs were identified, including 20:5n-3 (9.4%) and 22:6n-3 (7.3%) acids. Fourteen free sterols were present, including cholesterol at 31.3% of the sterol mixture and about 40% of phytosterols. These data on lipids of C. fornicata demonstrate their positive attributes for human nutrition and health. The PL mixture, rich in PC and polyunsaturated FAs, offers an interesting alternative source of high value-added marine lecithin.     

Identification of flea species using MALDI-TOF/MS

In the present study, a molecular proteomics (MALDI-TOF/MS) approach was used as a tool for identifying flea vectors. We measured the MS spectra from 38 flea specimens of 5 species including Ctenocephalides felis, Ctenocephalides canis, Archaeopsylla erinacei, Xenopsylla cheopis and Stenoponia tripectinata. A blind test performed with 24 specimens from species included in a library spectral database confirmed that MALDI-TOF/MS is an effective tool for discriminating flea species. Although fresh and 70% ethanol-conserved samples subjected to MALDI-TOF/MS in blind tests were correctly classified, only MS spectra of quality from fresh specimens were sufficient for accurate and significant identification. A cluster analysis highlighted that the MALDI Biotyper can be used for studying the phylogeny of fleas.     

Staphylococcus aureus from 152 cases of bovine,ovine and caprine mastitis investigated by Multiple-locus variable number of tandem repeat analysis (MLVA)

Staphylococcus aureus is one of the main etiological agents of mastitis in ruminants. In the present retrospective study, we evaluated the potential interest of a previously described automated multiple loci Variable Number of Tandem Repeats (VNTR) Assay (MLVA) comprising 16 loci as a first line tool to investigate the population structure of S. aureus from mastitis. We determined the genetic diversity of S. aureus strains from cases of clinical and subclinical mastitis in dairy cattle (n = 118, of which 16 were methicillin-resistant), sheep (n = 18) and goats (n = 16). The 152 strains could be subdivided into 115 MLVA genotypes (including 14 genotypes for the ovine strains and 15 genotypes for the caprine strains). This corresponds to a discriminatory index (D) value of 0.9936. Comparison with published MLVA data obtained using the same protocol applied to strains from diverse human and animal origins revealed a low number (8.5%) of human-related MLVA genotypes among the present collection. Eighteen percent of the S. aureus mastitis collection belonged to clonal complexes apparently not associated with other pathological conditions. Some of them displayed a relatively low level of diversity in agreement with a restricted ecological niche. These findings provide arguments suggesting that specific S. aureus lineages particularly adapted to ruminant mammary glands have emerged and that MLVA is a convenient tool to provide a broad overview of the population, owing to the availability via internet of databases compiling published MLVA genotypes.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0097-4) contains supplementary material, which is available to authorized users.     

Effects of Soil Type and Tillage on Protein and Starch Quality in Three Related Wheat Genotypes

An experiment was conducted using three related wheat (Triticum aestivum L.) genotypes grown on two different soil types (a grey vertosol and a red kandosol) using two different tillage practices (complete and zero) to test the effects of environmental influence on grain quality. Wheat grains from plants grown in each environment were milled into flour; protein and starch were analyzed. The soil type had the biggest impact on both protein and starch content, with the grains from the grey vertosol soil having higher total, insoluble, and soluble protein contents, and lower starch content and flour swelling values. When protein was analyzed using SDS‐PAGE, the major difference observed between grains from the two soil types was an increased intensity of polypeptide bands corresponding to β‐amylase in grains from plants grown on grey vertosol soils.     

Modifications of trout (Oncorhynchus mykiss) muscle proteins by preslaughter activity

The effect of two different preslaughter procedures (limited or 15-min intense muscular activity) on muscle trout proteins was investigated. Muscle was sampled 45 min and 24 h post-mortem, proteins were separated using two-dimensional electrophoresis, and spots of interest were tentatively identified by MALDI-TOF spectrometry. Twenty-nine and 4 spots were differentially represented between the two groups of fish at 45 min and 24 h post-mortem, respectively. Spots that could be identified corresponded mainly to proteins involved in energy-producing pathways (triosephosphate isomerase, enolase, pyruvate dehydrogenase) or to structural proteins (desmin, cap-Z, myosin heavy chain fragment). Persistent under-representation of desmin, a key cytoskeletal protein, in fish submitted to intense muscular activity suggests that such a preslaughter treatment can have an effect on post-mortem muscle integrity.     

Adenylate cyclases of Trypanosoma brucei inhibit the innate immune response of the host

The parasite Trypanosoma brucei possesses a large family of transmembrane receptor-like adenylate cyclases. Activation of these enzymes requires the dimerization of the catalytic domain and typically occurs under stress. Using a dominant-negative strategy, we found that reducing adenylate cyclase activity by about 50% allowed trypanosome growth but reduced the parasite's ability to control the early innate immune defense of the host. Specifically, activation of trypanosome adenylate cyclase resulting from parasite phagocytosis by liver myeloid cells inhibited the synthesis of the trypanosome-controlling cytokine tumor necrosis factor-α through activation of protein kinase A in these cells. Thus, adenylate cyclase activity of lyzed trypanosomes favors early host colonization by live parasites. The role of adenylate cyclases at the host-parasite interface could explain the expansion and polymorphism of this gene family.     

Local expansion and selection of soil bacteria in a glacier forefield

In the environment of Alpine glacier forefields, bacteria represent the functional link between initial soil development and secondary colonization. An important question is whether soil bacteria are adapted to the poor nutrient availability in this environment and whether selection processes significantly limit their local expansion along the gradients of chemical and biological factors. To address these questions, we used a two‐dimensional sampling strategy at our study site, the Damma glacier forefield, Switzerland, to characterize the soil environment by selected biogeochemical parameters. Our results revealed two contrasting sites that differed in time since deglaciation (‘young’ and ‘old’ soil, ≤ 13 years and > 53 years of ice‐free conditions) and vegetation cover (threefold increase), but also showed significant differences in soil properties thought to be relevant for the successional development of glacier forefield soils, such as pH (0.5‐fold decrease), ammonium (0.5‐fold decrease) concentration, and the content of total organic carbon (10‐fold increase). Isolated strains, affiliated to the genus Pseudomonas, according to their 16S rDNA profile, were tested experimentally for local adaptation by a reciprocal transfer experiment. The bacteria from both young and old soils showed a significant interaction with their local environment; the direction of the interaction, however, did not follow the expected pattern of local adaptation. The significant genoype‐with‐environment interaction indicates a limited local expansion of soil bacteria in the Damma glacier forefield over the range of only 110 m. The lack of local adaptation could be explained by environmental constraints such as exclusion of competition, different migration behaviour of bacteria from young and old soil, or the ability of early colonizing bacteria to survive under the glacier ice.