Induction of Defense Gene Expression and the Resistance of Date Palm to Fusarium oxysporum f. sp. Albedinis in Response to Alginate Extracted from Bifurcaria bifurcata

In many African countries, the Bayoud is a common disease spread involving the fungus Fusarium oxusporum f. sp. albedinis (Foa). The induction of plant natural defenses through the use of seaweed polysaccharides to help plants against pathogens is currently a biological and ecological approach that is gaining more and more importance. In the present study, we used alginate, a natural polysaccharide extracted from a brown algae Bifurcaria bifurcata, to activate date palm defenses, which involve phenylalanine ammonia-lyase (PAL), a key enzyme of phenylpropanoid metabolism. The results obtained showed that at low concentration (1 g·L⁻¹), alginate stimulated PAL activity in date palm roots 5 times more compared to the negative control (water-treated) after 24 h following treatment and 2.5 times more compared to the laminarin used as a positive stimulator of plant natural defenses (positive control of induction). Using qRT-PCR, the expression of a selection of genes involved in three different levels of defense mechanisms known to be involved in response to biotic stresses were investigated. The results showed that, generally, the PAL gene tested and the genes encoding enzymes involved in early oxidative events (SOD and LOX) were overexpressed in the alginate-treated plants compared to their levels in the positive and negative controls. POD and PR protein genes selected encoding β-(1,3)-glucanases and chitinases in this study did not show any significant difference between treatments; suggesting that other genes encoding POD and PR proteins that were not selected may be involved. After 17 weeks following the inoculation of the plants with the pathogen Foa, treatment with alginate reduced the mortality rate by up to 80% compared to the rate in control plants (non-elicited) and plants pretreated with laminarin, which agrees with the induction of defense gene expression and the stimulation of natural defenses in date palm with alginate after 24 h. These results open promising prospects for the use of alginate in agriculture as an inducer that triggers immunity of plants against telluric pathogens in general and of date palm against Fusarium oxysporum f. sp. albedinis in particular.     

Evidence of diadromy in the French Polynesian Kuhlia malo (Teleostei: Percoidei) inferred from otolith microchemistry analysis

Abstract – Strontium/Calcium (Sr/Ca) and Barium/Calcium (Ba/Ca) ratios were measured in 60 otoliths of the French Polynesian flagtail Kuhlia malo. Both elemental ratios were needed to correctly distinguish residence in marine, brackish and fresh water. High Sr/Ca and low Ba/Ca around the nucleus of all otoliths provided evidence of marine residency during the early life of all individuals. At about 0.5 mm from the core, Ba/Ca increased rapidly and remained high in all otoliths. High and unstable Sr/Ca ratios were attributed to estuarine residency, whereas low and stable Sr/Ca ratios were attributed to freshwater residency. These chemical signatures inferred that following recruitment to rivers, some fish remained in the estuary, while others moved upstream. Some individuals shared their time between fresh and brackish waters later in life. The findings of this study have important implications for the protection of nearshore, brackish and freshwater environments for the conservation of this French Polynesian diadromous species.     

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.     

Environmental correlates of body size distribution in Cyprinidae (Actinopterygians) depend on phylogenetic scale

The pattern of increasing species body size with increasing latitude has been noticed in different groups of animals. Here, we used seven key environmental factors and independent contrasts to assess body size latitudinal clines in Cyprinidae at two phylogenetic levels (inter‐ and intragenera), which were defined using a genus‐level supertree. Model selection procedures revealed that environmental factors shaping body size variation in Cyprinidae differed according to the phylogenetic scale considered. At the higher phylogenetic level, we found that both temperature (negative effect) and habitat availability (positive effect of drainage basin surface area) constituted mechanistic explanations of large‐scale body size distribution. No temperature‐related body size cline was observed at the intragenus level. Instead, competitive interaction (negative effect of species richness), habitat availability (positive effect of drainage basin surface area), migration ability and available energy (positive effects of glacial coverage and actual evapotranspiration) constitute alternative explanations at this lower phylogenetic scale. We conclude that (i) at the intergenus level, cyprinids do show a tendency to be smaller at high temperatures and larger at low temperatures, (ii) this tendency no longer exists at the intragenus level, (iii) latitude per se is a weak predictor of body size clines whatever the taxonomic level analysed, (iv) generalising geographical body size patterns may be rendered difficult by the superimposition of a series of mechanisms across different taxonomic scales, and (v) habitat size, here acting positively at both taxonomic scales, may play a major role in shaping riverine species body size clines.     

Larval traits of the Caribbean amphidromous goby Sicydium punctatum (Gobioidei: Sicydiinae) in Guadeloupe

Amphidromous fish are the biggest contributor to the diversity of fish communities in river systems of Caribbean islands. Among them, Sicydium punctatum Perugia, 1896, which is endemic to the West Indies, represents the vast majority of fish in these rivers. The otolith microstructure and the biometry of S. punctatum postlarvae from Guadeloupe were investigated to explore the dispersal modalities of this species through an appreciation of the growth pattern, the pelagic larval duration (PLD) and the size‐at‐recruitment. The study was made on one cohort of 83 recruited postlarvae, fished at the Capesterre River's mouth on 2 November 2011. The mean (±SD) size‐at‐recruitment of the postlarvae was 24.6 ± 1.3 mm (range of 20.5–28.1 mm, n = 83). We found a mean (±SD) PLD of 72.2 ± 10.5 days (range of 54–101.5 days, n = 67). The growth rate estimated from the otolith increments showed a globally decreasing pattern during the marine larval phase. Growth rates at the beginning of the larval stage were significantly different between hatching periods, suggesting a relation between the hatching period and the growth rate in S. punctatum. This study adds on to the general understanding of the life cycle of S. punctatum in Guadeloupe that will help implement strategies to manage amphidromous fish populations in the Caribbean region.     

Factors influencing the survivorship of the burrowing nematode,Radopholus similis (Cobb.) Thorne in two types of soil from banana plantations in Martinique

The burrowing nematode, Radopholus similis (Cobb.) Thorne, causes the most damage to bananas. To minimize nematicide applications, cropping systems that use fallow, crop rotation and clean planting material have been developed in the French West Indies. In order to optimize the benefit of the intercropping period, we studied the survivorship of R. similis in different soil types and conditions. We monitored the survivorship of calibrated populations of R. similis in the laboratory on a Nitisol and on an Andosol, two soils derived from volcanic ashes and pumices. We studied water potentials ranging from 0 to ?700 kPa on undisturbed soil and on soil previously frozen to get rid of living nematodes. Mortality of adult R. similis decreased regularly, and was fairly well described by Teissier's model. In the previously frozen soils, R. similis survived longer in wet soils (half-life of 21–46 days at 0 to ?5 kPa) than in dry soils (half-life of less than 10 days between ?80 and ?250 kPa). In contrast, in undisturbed soils, R. similis survived longer in dry soils: half-lives ranged from 57 days at ?273 kPa to 17 days at water saturation in the Andosol, and 36 days at ?660 kPa to 14 days at water saturation in the Nitisol. These results are consistent with the absence of anhydrobiosis in R. similis, unlike Pratylenchus coffeae. P. coffeae survivorship curves over time do not follow a model derived from exponential decrease like Teissier's model. These results also show that the recommended one year host-free period required to sanitize soils cannot be shortened without risk, even if flooding the soil could improve it.     

RAPD Characterization of Single Females of the Root-knot Nematodes, Meloidogyne Spp.

Random amplified polymorphic DNA fingerprinting was performed on single females of root-knot nematodes, Meloidogyne spp., using a new procedure for DNA isolation. One-fourth of the total DNA amount isolated from a single female proved to be sufficient as a template in a polymerase chain reaction. Electrophoretic patterns of the amplified fragments were reproducible between replicates from a single female or sister females from the same progeny, and identical to those obtained with genomic DNA purified from a large number of nematodes. Moreover, a comparative analysis over three successive generations showed stability of the amplification patterns, thus demonstrating the utility of this procedure for epidemiological and ecological studies on root-knot nematodes.     

A retrospective study of 30 cases of frozen lamellar corneal graft in dogs and cats

Frozen lamellar corneal grafts and nictitating membrane flaps were used in 18 dogs and 12 cats to repair deep corneal defects. In all dogs either melting corneal ulcers or descemetoceles were present. In the 12 cats, nine had either a melting corneal ulcer or descemetocele, two animals had acute bullous keratopathy, and one cat had corneal sequestrum. Initial vascularization with gradual clearing of the graft occurred during the first 45 days postoperatively. At 60 days postoperatively, all eyes were visual. Frequent postoperative complications included: focal dehiscence of the wound ( n  = 9); melting of part of the graft ( n  = 7); and pigmentation of the graft ( n  = 4). The frozen lamellar corneal graft was a very safe technique, and restored the tectonic and the optical function of the cornea. It provided the best results in corneas with nonperforating corneal defects. This technique provides poorer results when the cornea was perforated prior to surgery or during the surgical procedure.     

Comparison of genome segments 2, 7 and 10 of bluetongue viruses serotype 2 for differentiation between field isolates and the vaccine strain

Bluetongue (BT) virus serotype 2 (BTV 2) was first confirmed in Tunisia in February 2000 and has since spread northward and westward, infecting several other countries and islands, including Corsica, where clinical disease was reported in October 2000. BT was again reported on the Island in July 2001, some six months after a vaccination campaign against BTV 2. The molecular relationship between isolates of the BTV 2 Corsican wild-type viruses from 2000 and 2001, and the attenuated BTV 2 vaccine were determined by comparing corresponding sequences of genome segments 2, 7 and 10 with each other and with already published sequences available in the genome database. Complete genetic stability was observed between the isolates of the Corsican BTV 2. There was some divergence between the nucleotide sequences of segment 10 obtained from the wild-type and vaccine virus strains. Based on these differences, primers were selected that could be used in RT-PCR to differentiate between the wild-type and the vaccine viruses.     

Characterization of the Exoglucanase-Encoding Gene PaEXG2 and Study of Its Role in the Biocontrol Activity of Pichia anomala Strain K

ABSTRACT The PaEXG2 gene, encoding an exo-beta-1,3-glucanase, was isolated from the biocontrol agent Pichia anomala strain K. PaEXG2 has the capacity for coding an acidic protein of 427 amino acids with a predicted molecular weight of 45.7 kDa, a calculated pI of 4.7, and one potential N-glycosylation site. PaEXG2 was disrupted by the insertion of the URA3 marker gene, encoding orotidine monophosphate decarboxylase in strain KU1, a uracil auxotroph derived from strain K. Strain KU1 showed inferior biocontrol activity and colonization of wounds on apples, compared to the prototrophic strain. Antagonism and colonization were recovered after the restoration of prototrophy by transformation with the URA3 gene. Integrative transformation was shown to be mostly ectopic in strain K descendants (only 4% of integration by homologous recombination). PaEXG2 disruption abolished all detectable extracellular exo-beta-1,3-glucanase activity in vitro and in situ but did not affect biocontrol of Botrytis cinerea on wounded apples.