A 13CO2 Enrichment Experiment to Study the Synthesis Pathways of Polyunsaturated Fatty Acids of the Haptophyte Tisochrysis lutea

The production of polyunsaturated fatty acids (PUFA) in Tisochrysis lutea was studied using the gradual incorporation of a ¹³C-enriched isotopic marker, ¹³CO₂, for 24 h during the exponential growth of the algae. The ¹³C enrichment of eleven fatty acids was followed to understand the synthetic pathways the most likely to form the essential polyunsaturated fatty acids 20:5n-3 (EPA) and 22:6n-3 (DHA) in T. lutea. The fatty acids 16:0, 18:1n-9 + 18:3n-3, 18:2n-6, and 22:5n-6 were the most enriched in ¹³C. On the contrary, 18:4n-3 and 18:5n-3 were the least enriched in ¹³C after long chain polyunsaturated fatty acids such as 20:5n-3 or 22:5n-3. The algae appeared to use different routes in parallel to form its polyunsaturated fatty acids. The use of the PKS pathway was hypothesized for polyunsaturated fatty acids with n-6 configuration (such as 22:5n-6) but might also exist for n-3 PUFA (especially 20:5n-3). With regard to the conventional n-3 PUFA pathway, Δ6 desaturation of 18:3n-3 appeared to be the most limiting step for T. lutea, “stopping” at the synthesis of 18:4n-3 and 18:5n-3. These two fatty acids were hypothesized to not undergo any further reaction of elongation and desaturation after being formed and were therefore considered “end-products”. To circumvent this limiting synthetic route, Tisochrysis lutea seemed to have developed an alternative route via Δ8 desaturation to produce longer chain fatty acids such as 20:5n-3 and 22:5n-3. 22:6n-3 presented a lower enrichment and appeared to be produced by a combination of different pathways: the conventional n-3 PUFA pathway by desaturation of 22:5n-3, the alternative route of ω-3 desaturase using 22:5n-6 as precursor, and possibly the PKS pathway. In this study, PKS synthesis looked particularly effective for producing long chain polyunsaturated fatty acids. The rate of enrichment of these compounds hypothetically synthesized by PKS is remarkably fast, making undetectable the ¹³C incorporation into their precursors. Finally, we identified a protein cluster gathering PKS sequences of proteins that are hypothesized allowing n-3 PUFA synthesis.     

Clinical observations on changes in concentrations of hormones in plasma of two stallions with thermally-induced testicular degeneration

To investigate effects of thermally-induced testicular degeneration on hormonal and seminal parameters in stallions, the scrotum was insulated for 36 hours in two mature (5-year-old mixed breed and 11-year-old Throughbred) stallions. Semen was collected daily for 10 days (DSO) prior to, and at intervals after, scrotal insulation. When DSO determinations were not being made, semen was collected 3 times weekly. Jugular blood samples were collected at 15-minute intervals for 6 hours from each stallion prior to, and at intervals after, scrotal insulation. A mouse interstitial cell testosterone assay was modified to quantify biologic activity of equine luteinizing hormone (BLH) in plasma samples. Immunoactive luteinizing hormone (ILH) and testosterone (T) concentrations were determined in plasma samples by routine RIA procedures. Percentages of progressively motile and morphologically normal spermatozoa began to decrease by 1 to 2 weeks postinsulation, reached nadir values at 3 to 3-1/2 weeks postinsulation, and returned to preinsulation values by 7 weeks postinsulation. Total number of spermatozoa and total number of progressively motile, morphologically normal spermatozoa in ejaculates at DSO returned to normal by 8 weeks postinsulation in stallion 2 and 12 weeks postinsulation in stallion 1. Concentrations of BLH and ILH increased, and while T concentrations decreased, immediately postinsulation. The increase in ILH concentrations was greater than the increase in BLH concentrations, resulting in a decrease in the BLH:ILH (B:I) ratio. Following the peak in LH secretion immediately postinsulation, LH concentrations gradually decreased while T concentrations increased. The B:I ratio was elevated from 1 to 13 weeks postinsulation compared to immediately postinsulation. In addition to changes in spermatozoal quality in ejaculates, stallion response to scrotal insulation included increased secretion of luteinizing hormone and impaired Leydig cell function (as determined by reduced testosterone concentration in circulating plasma). The proportion of biologically active LH secreted in response to thermal testicular injury increased during the recovery phase.     

The effect of different HUFA enrichment emulsions on the nutritional value of rotifers (Brachionus plicatilis) fed to larval haddock (Melanogrammus aeglefinus).

A feeding study was conducted in the winter 2001 to determine the effects of feeding rotifers (Brachionus plicatilis) enriched with various levels of essential fatty acids on the growth and survival of haddock larvae (Melanogrammus aeglefinus). Rotifer enrichment treatments were: 1) mixed algae, 2) high DHA (docosahexaenoic acid, 22:6n-3), 3) high DHA and EPA (eicosapentaenoic acid, 20:5n-3), and 4) DHA, EPA, and AA (arachidonic acid, 20:4n-6). Larvae were fed rotifers enriched with the different treatments from days 1 to 16 post-hatch. From day 17 until 25 all treatment groups were fed rotifers reared on mixed algae and then weaned onto the International Council for Exploration of the SEA (ICES) Standard Reference Weaning diet (http://allserv.rug.ac.t/aquaculture/rend/rend.htm) over a five day period. The experiment was terminated on day 41 post-hatch. The enrichment treatments affected the fatty acid composition of the rotifers and correlated with the accumulation of these fatty acids in the haddock larvae. However, no significant differences in larval growth or survival to 40 days post hatch were detected, suggesting that all treatments provided the minimal essential fatty acid requirements for haddock.     

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.     

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.