Improved mating efficiency inAcetabularia acetabulum: recovery of 48–100% of the expected zygotes

Summary We have improved zygote recovery 11–1,000 fold by optimizing the physiology of gamete release and mating inAcetabularia acetabulum. Gamete release was affected by agar purity, concentration, and volume/gametangial pair. Cold pre-treatment of gametangia (14–30 d at 10°C in the dark) synchronized subsequent gamete release at 21°C in the light. Cold pre-treatment was nearly twice as effective in synchronizing subsequent gamete release when intact, gametangia-bearing caps rather than isolated gametangia were pretreated. Synchronizing gamete release doubled mating efficiency. In a wild-type laboratory strain ofA. acetabulum, there were 1,561±207 gametes/gametangium which had half-lives of 14.5 d in 0.1% seawater-agar. We recovered 48–93% of the expected numbers of zygotes from a mass mating of 8 to 1,226 gametangia and 11–128% of the expected numbers of zygotes from mating single gametangial pairs: the large range in the calculated mating efficiency may be attributable to the variation in the numbers of gametes made per gametangium. Zygote recovery from single gametangial pairs was highly dependent on the volume of mating matrix. In addition, most zygotes recovered were unattached to any other zygotes in the subsequent generation (> 95% single cells from matings of 1–500 gametangial pairs). Our improvements in mating conditions and zygote recovery (1) have facilitated cell manipulation and culture ofA. acetabulum in the laboratory; and (2) have made controlled crosses for selection and genetic analysis of mutants feasible. These advances have removed a major barrier to genetic analysis of development inAcetabularia.Abbreviations LB Luria-Bertani bacteriological broth - SE standard error of the mean - T_g agar gelling temperatures - DAPI 4,6-diamidino-2-phenylindole     

The ant Lasius niger shows no relationship between task efficiency and body size variation among workers

In ants, workers of different sizes may perform various tasks, even in so-called monomorphic species with relatively low body size variation. However, it is unclear if the body size diversity of monomorphic workers correlates with task efficiency, especially in stressful contingencies. Here we tested if the body size variation of workers corresponds with its efficiency in transferring pupae. Transferring brood is a pre-set behavioral response to stress, e.g. suboptimal temperature. Here we applied a laboratory experiment simulating nest damage. The study was performed on the common garden ant (Lasius niger (Linnaeus, 1758)) – a species with no distinct worker subcastes. The efficiency of workers was measured as the latency of transferring pupae from a lit part of the experimental colony to a darkened part, while the body size diversity was expressed as the within-colony coefficient of variation in head width. We did not find any significant correlation between efficiency and body size variation. Summarizing the existing studies and the present results, we propose the hypothesis that the body size diversity of L. niger may have implications for workers’ division of labor but not for their task efficiency in a stressful contingency.     

The efficiency and effectiveness of different sea urchin removal methods for kelp forest restoration

Sea urchin overgrazing has caused widespread phase shifts from kelp forests to “urchin barrens” on many temperate reefs, reducing habitat complexity, productivity, and biodiversity. Sea urchin removal is increasingly used for kelp restoration; however, few studies have quantified the efficiency and effectiveness of different removal methods, resulting in limited understanding of their practicality. In this study, the efficiency (removal rate) and effectiveness (proportion removed) of four removal methods were evaluated in northeastern New Zealand. We compared culling or collecting sea urchins by either SCUBA or freediving in 128 small-scale plots (25 m²). We also evaluated the efficiency and effectiveness of culling in four large (1.6–2 ha) barren areas, scales relevant for restoration. On average, culling sea urchins was 1.9–4.4 times faster than collecting, and SCUBA was 1.5–3.3 times faster than freediving. Removal rates increased with sea urchin density, especially for culling on SCUBA, while freediving removal rates increased with experience. Effectiveness was lower in large-scale removals (86–93% of sea urchins ≥40 mm removed) compared to small-scale removals (98–99%), but sufficient for restoration objectives. Estimated time per area (using SCUBA culling) was similar across large-scale removals (49–57 hours/ha), despite an almost 2-fold variation in initial sea urchin densities (approximately 4–8 urchins/m²), suggesting area may better predict total removal time than simply number of sea urchins across low-density ranges. While sea urchin removal provides a rapid, feasible, and effective approach to restoring kelp in urchin barrens, restoration plans need to also address the causes of sea urchin overpopulation to ensure long-term benefits.     

Leptocephalus eel larvae will feed in aquaria

Synopsis Premetamorphosing larvae of Muraenesox cinereus (Muraenesocidae) and Conger myriaster (Congridae), caught and kept alive in aquaria, repeatedly bit pieces out of a lump of squid paste. By coloring the paste with Brilliant Red, ingestion and defecation were clearly seen through the transparent gut. The paste is not their natural food, but nevertheless a possible food item for rearing eels from eggs; most previous efforts have failed inasmuch as the larvae have starved.     

Dietary exposure and risk assessment to trace elements and iodine in seaweeds

IntroductionSeaweeds are a rich source of elements such as iodine, and are also able to accumulate contaminants such as trace elements.MethodsThe aim of this study was to assess the dietary exposure as well as the risk from iodine and trace elements in edible seaweeds for the French population using current consumption data. The contribution of seaweeds to overall dietary exposure to trace elements and iodine was evaluated, and for those substances with minimal contribution to overall dietary exposure, simulations were performed to propose increased maximal limits in seaweeds.ResultsCadmium, inorganic arsenic and mercury in seaweeds were very low contributors to total dietary exposure to these contaminants (0.7 % 1.1 % and 0.1 % on average, respectively). Dietary exposure to lead via seaweed may contribute up to 3.1 % of total dietary exposure. Dietary consumption of iodine via seaweed may contribute up to 33 % of total exposure to iodine, which makes seaweeds the strongest contributor to iodine in diet.DiscussionNew maximal values in seaweeds are proposed for the very low contributors to total dietary exposure: 1 mg/kg dw for cadmium, 10 mg/kg dw for inorganic arsenic and 0.3 mg/kg dw for mercury.     

Multikink quantile regression for longitudinal data with application to progesterone data analysis

Motivated by investigating the relationship between progesterone and the days in a menstrual cycle in a longitudinal study, we propose a multikink quantile regression model for longitudinal data analysis. It relaxes the linearity condition and assumes different regression forms in different regions of the domain of the threshold covariate. In this paper, we first propose a multikink quantile regression for longitudinal data. Two estimation procedures are proposed to estimate the regression coefficients and the kink points locations: one is a computationally efficient profile estimator under the working independence framework while the other one considers the within-subject correlations by using the unbiased generalized estimation equation approach. The selection consistency of the number of kink points and the asymptotic normality of two proposed estimators are established. Second, we construct a rank score test based on partial subgradients for the existence of the kink effect in longitudinal studies. Both the null distribution and the local alternative distribution of the test statistic have been derived. Simulation studies show that the proposed methods have excellent finite sample performance. In the application to the longitudinal progesterone data, we identify two kink points in the progesterone curves over different quantiles and observe that the progesterone level remains stable before the day of ovulation, then increases quickly in 5 to 6 days after ovulation and then changes to stable again or drops slightly.     

Effects of alginates on the ingestion and excretion of cholesterol in the rat

The effects of three forms of alginate (free acid, sodium and calcium salts) on ingestion and excretion of cholesterol in the rat were studied. For two weeks, Male Sprague-Dawley rats were fed a cholesterol-rich diet containing 3% alginate from the brown alga Ascophyllum nodosum. The food efficiency of the three types of alginate was: Na-alginate > Ca-alginate > alginic acid. A significant increase in the weight of cecum was also observed in alginate diets. Alginate was not effective in preventing the elevation of serum total cholesterol levels, although irregularly changing patterns were observed. The cholesterol level of liver showed a tendency to decrease with alginate feeding, while fecal excretion of cholesterol increased.     

Nutritional properties of four marine microalgae for albino rats

The nutritive value of the marine microalgaeTetraselmis suecica, Isochrysis galbana, Dunaliella tertiolecta andChlorella stigmatophora was studied in diets given to rats. Biological assays were carried out in order to determine the Protein Efficiency Ratio (PER) and the Food Conversion Efficiency (FCE). Each dried microalga was fed to weaning Wistar albino rats as the sole protein source at a protein level of 12%. Control rats were given diets containing 12 % casein. Food consumption was similar in all groups. PER values obtained were 1.14 withT. suecica diet, 1.13 withI. galbana diet, 2.07 withD. tertiolecta diet and 1.13 withC. stigmatophora diet (casein, 2.50). FCE values followed a similar pattern. The data showed that the marine microalgaD. tertiolecta is a source of protein of good quality. Its PER is quite high, compared to vegetable and cereal proteins, and compares favourably with other microbial protein sources, such as yeasts or different freshwater microalgae. Haematological tests showed no significant differences among the groups in haemoglobin levels, red and white blood cell counts, differential count and mean corpuscular volume. Different blood parameters were also determined and a significant decrease in triglycerides levels appeared with all the microalgal diets, whereas a significant decrease in cholesterol appeared inD. tertiolecta andC. stigmatophora diets.     

Effect of nitrogen application on growth and photosynthetic nitrogen use efficiency in two ecotypes of wild strawberry, Fragaria chiloensis

The relationships between increasing nitrogen fertilization and growth, maximum CO₂ assimilation and the initial slope of the CO₂ response curve were studied in 2 ecotypes of wild strawberry, Fragaria chiloensis (L.) Duchn. Nitrogen accumulation of CA11, an ecotype from a low-nutrient dune site, was greater at all nitrogen concentrations than that of RCP37, an ecotype from a higher-nutrient strand site. Maximum CO₂ assimilation, total Rubisco activity, dry weight, and initiation of leaves and crowns were higher in CAI1 than RCP37 as nitrogen treatment was increased from 0 to 200 mg ^l-1, whereas these parameters were lower in CAl1 when fertilized at 300 mg T¹, but not in RCP37. The mean leaf area of CA11 was greater than RCP37 when grown with no supplemental nitrogen, but mean leaf area of the 2 lines was similar under nitrogen fertilization. Maximum CO₂ assimilation and carboxylation efficiency increased with increasing leaf nitrogen in both clones. At equivalent concentrations of leaf nitrogen, RCP37 had higher CO₂ assimilation and carboxylation efficiency than CA11 and the difference between the 2 clones increased as ieaf nitrogen increased. Thus, RCP37 had a higher photosynthetic nitrogen use efficiency than CA11. However, at a given applied nitrogen level, CA11 allocated more nitrogen to a unit of leaf area so that photosynthetic rates were higher than RCP37, except at the highest application of 300 mg ^l-1. The high nitrogen accumulation capacity and resource allocation to fruiting structures (crowns) in CA11 leads us to suggest that this clone may possess genes that could increase fruit yield in cultivated strawberry.     

CO2 uptake by the cultivated hemiepiphytic cactus, Hylocereus undatus

The climate of the native tropical forest habitats of Hylocereus undatus, a hemiepiphytic cactus cultivated in 20 countries for its fruit, can help explain the response of its net CO2 uptake to environmental factors. Under wet conditions, about 85% of the total daily net CO₂ uptake occurs at night via Crassulacean acid metabolism, leading to a high water‐use efficiency. Total daily net CO₂ uptake is reduced 57% by only 10 days of drought, possibly involving stomatal closure induced by abscisic acid produced in the roots, which typically occupy a small substrate volume. Total daily net CO₂ uptake for H. undatus is maximal at day/night air temperatures of 30/20°C, optimal temperatures that are higher than those for desert cacti but representative of ambient temperatures in the tropics; its total daily net CO₂ uptake becomes zero at day/night air temperatures of 42/32°C. Stem damage occurs at 45°C for H. undatus, whose photosynthetic cells show little acclimation to high temperatures compared with other cacti and are also sensitive to low temperatures, ‐1.5°C killing half of these cells. Consistent with its shaded habitat, total daily net CO2 uptake is appreciable at a total daily PPF of only 2 mol m2 day' and is maximal at 20 mol m^?2 day^?1, above which photoinhibition reduces net CO₂ uptake. Net CO₂ uptake ability, which is highly correlated with stem nitrogen and chlorophyll contents, changes only gradually (halftimes of 2–3 months) as the concentration of applied N is changed. Doubling the atmospheric CO₂ concentration raises the total daily net CO₂ uptake of H. undatus by 34% under optimal conditions and by even larger percentages under adverse environmental conditions.