Multilocus sequence typing detects new Piscirickettsia salmonis hybrid genogroup in Chilean fish farms: Evidence for genetic diversity and population structure

Piscirickettsia salmonisis the causative bacterial pathogen of piscirickettsiosis, a salmonid disease that causes notable mortalities in the worldwide aquaculture industry. Published research describes the phenotypic traits, virulence factors, pathogenicity and antibiotic‐resistance potential for various P. salmonisstrains. However, evolutionary and genetic information is scarce for P. salmonis. The present study used multilocus sequence typing (MLST) to gain insight into the population structure and evolution of P. salmonis. Forty‐two Chilean P. salmonisisolates, as well as the type strain LF‐89^T, were recovered from diseased Salmo salar, Oncorhynchus kisutchand Oncorhynchus mykissfrom two Chilean Regions. MLST assessed the loci sequences of dnaK, efp, fumC, glyA, murG, rpoD and trpB. Bioinformatics analyses established the genetic diversity among P. salmonis isolates (H = 0.5810). A total of 23 sequence types (ST) were identified, 53.48% of which were represented by ST1, ST5 and ST2. Population structure analysis through polymorphism patterns showed few polymorphic sites (218 nucleotides from 4,010 bp), while dN/dS ratio analysis indicated purifying selection for dnaK, epf, fumC, murG, and rpoD but neutral selection for the trpB loci. The standardized index of association indicated strong linkage disequilibrium, suggesting clonal population structure. However, recombination events were detected in a group of seven isolates. Findings included genogroups homologous to the LF‐89^T and EM‐90 strains, as well as a seven‐isolate hybrid genogroup recovered from both assessed regions (three O. mykiss and four S. salar isolates). The presented MLST scheme has comparative potential, with promising applications in studying distinct P. salmonis isolates (e.g., from different hosts, farms, geographical areas) and in understanding the epidemiology of this pathogen.     

Fine‐scale habitat partitioning of Chilean and Peale's dolphins and their overlap with aquaculture

1. Predictive species distribution models (SDMs) have become powerful tools to determine habitat use patterns of mobile marine predators and their spatial overlap with potentially impacting anthropogenic activities.
2. This study used SDMs to investigate fine‐scale habitat use patterns of two poorly known and broadly sympatric coastal delphinids, Chilean dolphins (Cephalorhynchus eutropia) and Peale's dolphins (Lagenorhynchus australis), and their spatial interactions with intense aquaculture farming activities in the Chiloé archipelago, southern Chile.
3. A long‐term dataset (2002–2012) of boat‐based dolphin sightings and concurrently in situ collected environmental and anthropogenic variables was analysed using binomial Generalized Additive Models to investigate ecological drivers of each species' fine‐scale distribution and to predict dolphin occurrence spatially.
4. Chilean dolphins preferred shallow (<30 m deep), turbid waters, close to shore (<500 m) and river mouths which often placed them in sheltered bays and channels used intensively by shellfish farms. Peale's dolphins were also found in shallow waters but occurred over a wider range of conditions along more open or exposed coastlines. Both species had to navigate extensive salmon and shellfish farming sites to transit between areas of important habitat.
5. Sightings and predicted occurrence maps showed a clear pattern of spatial habitat partitioning between species, which remained stable across the 11 year study period. The identification of important habitat for Chilean dolphins warrants the consideration of spatially explicit conservation measures to limit the potential effects of overlapping salmon and shellfish farming.
6. The observed differences in ecological plasticity of the two sympatric species should be considered when evaluating and mitigating the effects of environmental change and ongoing anthropogenic pressures on their nearshore habitat. The estimated species–environment relationships could also be used to predict where dolphin habitat and anthropogenic activities are most likely to overlap in other parts of the species' ranges.

     

Astaxanthin from Haematococcus pluvialis Prevents Oxidative Stress on Human Endothelial Cells without Toxicity

Astaxanthin, a powerful antioxidant, is a good candidate for the prevention of intracellular oxidative stress. The aim of the study was to compare the antioxidant activity of astaxanthin present in two natural extracts from Haematococcus pluvialis, a microalgae strain, with that of synthetic astaxanthin. Natural extracts were obtained either by solvent or supercritical extraction methods. UV, HPLC-DAD and (HPLC-(atmospheric pressure chemical ionization (APCI)+)/ion trap-MS) characterizations of both natural extracts showed similar compositions of carotenoids, but different percentages in free astaxanthin and its ester derivatives. The Trolox equivalent antioxidant capacity (TEAC) assay showed that natural extracts containing esters displayed stronger antioxidant activities than free astaxanthin. Their antioxidant capacities to inhibit intracellular oxidative stress were then evaluated on HUVEC cells. The intracellular antioxidant activity in natural extracts was approximately 90-times higher than synthetic astaxanthin (5 µM). No modification, neither in the morphology nor in the viability, of vascular human cells was observed by in vitro biocompatibility study up to 10 µM astaxanthin concentrations. Therefore, these results revealed the therapeutic potential of the natural extracts in vascular human cell protection against oxidative stress without toxicity, which could be exploited in prevention and/or treatment of cardiovascular diseases.     

Bauhinia candicans stimulation of glucose uptake in isolated gastric glands of normal and diabetic rabbits

A butanol extract of dried leaves of Bahuinia candicans showed, in vitro, a stimulatory effect of glucose uptake in isolated gastric glands of normal and alloxan-diabetic rabbits. The extract (0.001-0.07 mg/mg protein) produced a dose-dependent effect and was similar to the effect of insulin (1-100 nM). These findings suggest that the extract contains compound(s) that efficiently increase glucose transport at the basolateral side of isolated gastric glands.     

Multivariate statistical analysis of mass spectra as a tool for the classification of the main humic substances according to their structural and conformational features

The aim of this work is to explore the suitability of the complementary use of mass spectra and the corresponding statistical analysis (principal components-Pareto analysis (PCA) and discriminant analysis (DA)) of these spectra to differentiate diverse humic samples as a function of their structural and conformational features. To this end, the mass spectra of humic samples belonging to the main humic fraction types (gray humic acid, brown humic acid, and fulvic acid) were obtained by electrospray ionization mass spectrometry (ESI-MS). The results obtained showed that the application of PCA yielded a clear separation between blanks and humic samples. However, a clear differentiation among the humic fraction types was not achieved. The DA of PCA data, however, yielded a clear separation among the humic substances (HS) samples belonging to each HS fraction type considered: gray humic acids, brown humic acids, and fulvic acids. These results showed that the mass spectra of each humic sample include characteristic mass/charge (m/z) distribution values that can be considered as a "fingerprint" representative of its specific structural features. Our results also indicate that, although the m/z values principally corresponded to single-charged ions, we cannot identify these molecular weight distributions with those of humic samples, since sample molecular fragmentation, as well as partial molecular ionization, cannot be ruled out under our experimental and instrumental conditions.     

Characterization of single‐nucleotide polymorphism markers in the Mediterranean mussel,Mytilus galloprovincialis

The Mediterranean mussel, Mytilus galloprovincialis, is one of the most important aquaculture species in Europe. Appropriate molecular markers are required to evaluate genetic resources and to trace genealogies in breeding programmes for improving mussel culture. Microsatellites have been commonly applied to this purpose in other species. However, Mediterranean mussel microsatellites have demonstrated high frequencies of null alleles that hamper accurate estimates of population parameters and confident parentage inferences. As alternative markers, we have characterized in silico 25 potential single‐nucleotide polymorphism (SNP) markers in the Mediterranean mussel from expressed sequence tag (EST) public databases. The genotyping of SNPs was performed using a single‐base extension approach. Their polymorphism was evaluated in 47 individuals from an Atlantic population. Out of the 25 potential SNPs tested, 12 were technically feasible (producing a single amplicon) and polymorphic. All were biallelic and had an unbiased heterozygosity ranging from 0.160 to 0.504. One SNP was from a mitochondrial gene. The combined potential of nuclear SNPs for parentage assignment gave an exclusion probability of a false couple of parents of 0.9471. These markers will be useful for evaluating resources and tracing genealogies in genetic breeding programmes implemented to solve the main problems of mussel culture.     

Chemical composition of the Lippia origanoides essential oils and their antigenotoxicity against bleomycin-induced DNA damage

The present work evaluated the chemical composition of the essential oils (EO) obtained from Lippia origanoides and their DNA protective effect against bleomycin-induced genotoxicity. L. origanoides EO chemical composition was determined by gas chromatography–mass spectrometry (GC–MS). The major compounds of the L. origanoides EOs were thymol (34–58%) and carvacrol (26%). The antigenotoxic effects of the EOs, major compounds and standard compound (epigallocatechin gallate) were assayed in co-incubation procedures using the SOS chromotest in Escherichia coli. Both EOs and their major compounds protected bacterial cells against bleomycin-induced genotoxicity indicating that these two compounds were principally responsible for the antigenotoxicity detected in the oils. Thymol and carvacrol antigenotoxicity was lower than those observed with epigallocatechin gallate. The results were discussed in relation to the chemopreventive potential of L. origanoides EOs and their major components, carvacrol and thymol.