Behavioural differences underlie toxicity and predation variation in blooms of Prymnesium parvum

Much of the evolutionary ecology of toxic algal blooms (TABs) remains unclear, including the role of algal toxins in the adaptive ‘strategies’ of TAB-forming species. Most eukaryotic TABs are caused by mixotrophs that augment autotrophy with organic nutrient sources, including competing algae (intraguild predation). We leverage the standing diversity of TABs formed by the toxic, invasive mixotroph Prymnesium parvum to identify cell-level behaviours involved in toxin-assisted predation using direct observations as well as comparisons between genetically distinct low- and high-toxicity isolates. Our results suggest that P. parvum toxins are primarily delivered at close range and promote subsequent prey capture/consumption. Surprisingly, we find opposite chemotactic preferences for organic (prey-derived) and inorganic nutrients between differentially toxic isolates, respectively, suggesting behavioural integration of toxicity and phagotrophy. Variation in toxicity may, therefore, reflect broader phenotypic integration of key traits that ultimately contribute to the remarkable flexibility, diversity, and success of invasive populations.     

The effects of aeration on growth and toxicity of Prymnesium parvum grown with and without algal prey

We investigated the effects of aeration on growth and toxicity of the haptophyte Prymnesium parvum in the presence and absence of the algal prey Rhodomonas salina. Batch monocultures of P-limited P. parvum and N and P sufficient R. salina and mixed cultures of the two microalgae were grown with no, low (20) and high (100) ml min⁻¹ aeration for 18 days. Cell growth of P. parvum and R. salina and cell toxicity of P. parvum were studied over the experimental period. The highest specific growth rates of P. parvum were found at low aeration rates. R. salina in monocultures showed typical growth patterns, while R. salina numbers declined rapidly in the mixed cultures. Of the initial cell densities, 98–100% of the R. salina cells were lysed or ingested within 24 h of mixing with P. parvum cells. The maxima P. parvum biomasses were significantly higher in the mixed cultures than in the monocultures. Cell toxicity of P. parvum increased significantly in response to aeration rates and the highest levels were found in the high aeration condition. Availability of prey and resupply of inorganic nutrients decreased P. parvum cell toxicity. Our study suggests that P. parvum is tolerant and is able to grow over a broad range of aeration and associated turbulence effects though low aeration represents an optimal condition for growth. As P. parvum toxicity was higher in the high aeration treatment we suggest that the higher concentrations of oxygen cause more toxins to be produced, as these are oxygen rich compounds. We suggest that oxygenation and turbulence of surface waters caused by mixing may be involved in promoting high toxic P. parvum blooms in shallow lakes and coastal waters.     

Effect of competition on stable carbon isotope ratios of two tussock grass species

Summary Previous studies have shown that plant carbon isotope composition varies when plants experience differences in water and nutrient availability. However, none have addressed the effect of root interactions, including competition for these soil resources, on carbon isotope ratios. We studied the effect of interspecific root interactions on the productivity and carbon isotope ratios of two Great Basin tussock grass species (Agropyron desertorum and Pseudoroegneria spicata). We compared grasses grown in mixture with sagebrush (Artemisia tridentara) to grasses in similar mixtures but where root interactions with sagebrush were limited by fiberglass partitions. During both years of the study, tussocks growing in competition with sagebrush produced tissue with more negative ¹³C values than grasses experiencing limited root interaction with sagebrush. The magnitude of this difference (0.5 to 0.9%) is similar to that found in other studies when soil fertility and moisture availability were altered.     

Effect of Illumination on Ichthyotoxin in an Axenic Culture of Prymnesium parvum Carter

SYNOPSIS. Cultures of Prymnesium parvum subjected to constant illumination failed to produce ichthyotoxin. On the other hand cultures subjected to alternate periods of light and darkness showed a gradually rising ichthyotoxic activity during the dark period reaching a maximum after about 7 hours.     

Effect of lipid removal on carbon and nitrogen stable isotope ratios in crustacean tissues

The analysis of tissue's naturally occurring stable carbon and nitrogen isotope ratios is a useful tool to delineate trophic relationships. However, the interpretation of δ¹³C and δ¹⁵N is complicated by the influence of multiple factors such as the tissue-specific lipid content. The aim of this work was to evaluate the effects of lipid extraction on δ¹³C and δ¹⁵N compositions in muscle, hepatopancreas and gonads of a marine decapod crustacean, the spider crab Maja brachydactyla. Samples were analyzed for stable isotopes before and after lipid removal, using a derived Soxhlet extraction method. Differences in δ¹³C and δ¹⁵N were measured among tissues before and after treatment. Lipid extraction of muscle did not have a significant effect on either δ¹³C or δ¹⁵N. By contrast, ecologically significant shifts for both carbon and nitrogen stable isotopes ratios (+ 2.9 ± 0.8‰ for δ¹³C, and + 1.2 ± 0.7‰ for δ¹⁵N) were noticed in the hepatopancreas. In regard to gonads, lipid extraction led to a shift only on δ¹³C (+ 1.3 ± 0.3‰). Finally, the derived Soxhlet extraction method removed the lipid influence for δ¹³C, and had an effect on δ¹⁵N composition for lipid-rich samples. We recommend this treatment for carbon stable isotope studies on decapod crustacean lipid-rich tissues.     

预处理及保存方法对浮游动物碳、氮稳定同位素比值的影响

研究了不同预处理及保存方法对浮游动物碳、氮稳定同位素比值的影响.结果表明:未清养的舌状叶镖水蚤PhyHodiaptomus tunguidus与清养3h的相比δ¹³C和δ¹⁵N的差异均不显著,但肠道内含物造成了重复样品间的较大偏差;去除脂肪后的舌状叶镖水蚤δ¹³C和δ¹⁵N均显著升高;经甲醇保存后,舌状叶镖水蚤和奥氏秀体溞Diaphanosoma orghidani transamurensis的δ¹³C均有所升高,但不显著;经乙醇保存后,舌状叶镖水蚤δ¹³C显著升高,而奥氏秀体溞的δ¹³C也有所升高但不显著;经过甲醛保存后,舌状叶镖水蚤和奥氏秀体溞的δ¹³C降低,但不显著;冰冻保存使奥氏秀体溞的δ¹³C显著降低,而使舌状叶镖水蚤的δ¹³C也有所降低但不显著.四种保存方法对舌状叶镖水蚤和奥氏秀体溞的δ¹⁵N都没有显著影响.在进行不同研究间的比较时,必须考虑样品保存与处理方法造成的差异.同时,食物网结构的稳定同位素研究有待建立统一的样品保存与处理方法.     

Disentangling effects of growth and nutritional status on seabird stable isotope ratios

A growing number of studies suggest that an individual’s physiology affects its carbon and nitrogen stable isotope signatures, obscuring a signal often assumed to be only a reflection of diet and foraging location. We examined effects of growth and moderate food restriction on red blood cell (RBC) and feather δ¹⁵N and δ¹³C in rhinoceros auklet chicks (Cerorhinca monocerata), a piscivorous seabird. Chicks were reared in captivity and fed either control (75 g/day; n = 7) or ~40% restricted (40 g/day; n = 6) amounts of high quality forage fish. We quantified effects of growth on isotopic fractionation by comparing δ¹⁵N and δ¹³C in control chicks to those of captive, non-growing subadult auklets (n = 11) fed the same diet. To estimate natural levels of isotopic variation, we also collected blood from a random sample of free-living rhinoceros auklet adults and chicks in the Gulf of Alaska (n = 15 for each), as well as adult feather samples (n = 13). In the captive experiment, moderate food restriction caused significant depletion in δ¹⁵N of both RBCs and feathers in treatment chicks compared to control chicks. Growth also induced depletion in RBC δ¹⁵N, with chicks exhibiting lower δ¹⁵N when they were growing the fastest. As growth slowed, δ¹⁵N increased, resulting in an overall pattern of enrichment over the course of the nestling period. Combined effects of growth and restriction depleted δ¹⁵N in chick RBCs by 0.92‰. We propose that increased nitrogen-use efficiency is responsible for ¹⁵N depletion in both growing and food-restricted chicks. δ¹⁵N values in RBCs of free-ranging auklets fell within a range of only 1.03‰, while feather δ¹⁵N varied widely. Together, our captive and field results suggest that both growth and moderate food restriction can affect stable isotope ratios in an ecologically meaningful way in RBCs although not feathers due to greater natural variability in this tissue.     

Growth and Colony Formation of the Phytoflagellate Prymnesium parvum Carter on Solid Media

SYNOPSIS. Colonies of Prymnesium parvum from single cells were obtained in an atmosphere of high humidity, in soft-agar overlayers on solid medium. Light intensity above 60 foot candles at the surface of the agar plates inhibited colonial development. Such colonies were used to obtain pure lines of the phytoflagellate.
A method for direct demonstration of hemolysin formation on agar plates was developed. The occurrence of hemolytic activity by P. parvum colonies was light-dependent.     

Ecology and physiology of White River mammals based on stable isotope ratios of teeth

To characterize the ecology and physiology of common late Eocene–early Oligocene White River mammals, we analyzed the carbon and oxygen isotope composition of tooth enamel carbonate for six of the most abundant taxa: the perissodactyls Brontops (brontothere), Mesohippus (equid) and Subhyracodon (rhino); and the artiodactyls Merycoidodon (oreodont), Leptomeryx (leptomerycid) and Poebrotherium (camelid).δ¹³C values of middle and rear molars (M2s and M3s) and premolars range from ? 13.1‰ to ? 7.7‰ (V-PDB), consistent with pure C3 diets. In the late Eocene, Mesohippus, Merycoidodon, and Leptomeryx show indistinguishable average δ¹³C values (～ ? 10‰). In contrast, Brontops and Subhyracodon exhibit lower (? 11.2‰) and higher (? 8.7‰) values, respectively. Early Oligocene values for Mesohippus and Merycoidodon remain indistinguishable from each other and lower than the value of Leptomeryx, Poebrotherium, and Subhyracodon (～ ? 8.5‰). These results likely indicate niche separation in terms of habitat preference between the investigated sympatric taxa. More specifically, assuming a δ¹³C value of atmospheric CO₂ of ? 5.5‰, our data suggest a preference of Brontops for mesic forested areas, of Mesohippus and Merycoidodon for woodlands, and of Subhyracodon and Poebrotherium for more open habitats (e.g., grasslands). The higher Oligocene versus Eocene average δ¹³C exhibited by Leptomeryx possibly reflects a preference of the new Oligocene species L. evansi for more open and/or xeric habitats relative to the Eocene species L. speciosus.Late Eocene and early Oligocene average δ¹⁸O of Mesohippus, Merycoidodon, Leptomeryx, and Subhyracodon are similar (～ 25‰, V-SMOW), possibly indicating comparable water dependency for these taxa. In contrast, the higher δ¹⁸O of Poebrotherium (26.6‰) suggests lower water dependency whereas the lower δ¹⁸O of Brontops (23.0‰) may result from a high water dependency or, more likely, from its preference for humid habitats. Because hind-gut fermentation in perissodactyls requires high water intake, whereas fore-gut fermentation does not, our results might indicate the presence of fore-gut fermentation in early Oligocene camelids but its absence or incomplete development in late Eocene–early Oligocene oreodonts and leptomerycids.     

Preservation effects on stable isotope ratios and consequences for the reconstruction of energetic pathways

Stable isotope analysis provides a powerful tool for describing the energetic pathways in a variety of ecosystems. However, isotope ratios of animal tissues can be altered by preservation methods, potentially leading to biased estimates of energy pathways when they are not taken into account. Here, we investigated the direct preservation effects of formalin, ethanol, NaCl, and drying on the δ¹³C and δ¹⁵N of fish muscle tissues, as well as the ultimate effects on the reconstruction of the energy pathways. All preservation methods, except drying, had significant impacts on δ¹³C and δ¹⁵N values. The effects of preservation appear to be highly taxa-specific and no significant time-dependent variations in nearly 2-year duration of preservation. δ¹³C and δ¹⁵N values were generally changed dramatically within the early stage of the preservation process and became stable over a relatively long-term preservation. Using an isotopic balance mixing model, the isotope-based food web reconstruction reveals that, without preservation correction, the importance of the pelagic energetic pathways for the fishes could be misestimated, except for the drying preservation. These results highlight that preservation can bias the interpretation of food web reconstruction results.     

Global dataset for carbon and nitrogen stable isotope ratios of lotic periphyton

Carbon and nitrogen stable isotope ratios (δ¹³C and δ¹⁵N) have been widely employed in food web analysis. In lotic environments, periphyton is a major primary producer that makes a large contribution to food web production as well as carbon and nitrogen cycling. While the δ¹³C and δ¹⁵N values have many advantages as a natural tracer, the controls over their high spatial and temporal variability in stream periphyton are not well known. Here, we present the global dataset of δ¹³C and δ¹⁵N values of lotic periphyton from 54 published and two unpublished sources, including 978 observations from 148 streams/rivers in 38 regions around the world, from arctic to tropical sites. The 54 published sources were articles recorded during the period of 1994–2016 in 25 academic journals. The two unpublished sources were from the authors’ own data. The dataset showed that δ¹³C and δ¹⁵N values of periphyton ranged from ?47.3 to ?9.3‰ and from ?5.6 to + 22.6‰, respectively. The dataset also includes physicochemical factors (altitude, coordinates, catchment area, width, depth, geology, vegetation, canopy coverage, biome, season, presence of anadromous salmon, temperature, pH, current velocity, and discharge), nutrient data (nitrate and ammonium concentrations), and algal attributes (chlorophyll a concentration, algal species compositions, and carbonates removal) in streams/rivers studied, all of which may help interpret the δ¹³C and δ¹⁵N values of periphyton. The metadata file outlines structure of all the data and with references for data sources, providing a resource for future food web studies in stream and river ecosystems.     

Osteopenia and stable isotope ratios in bone collagen of Nubian female mummies

Stable carbon and nitrogen isotopes were analysed on bone collagen of 43 Sudanese Nubians from the X-Group period to test dietary hypotheses for the high frequency of osteopenia in this population. Stable carbon isotope ratios indicate that both normal and osteopenic individuals consumed the same mixed diet of C3 and C4 sources, which are assumed to have been constituted by the grain staples wheat/barley and sorghum/millet respectively. Females with osteopenia, however, have significantly elevated δ¹⁵N values. The enrichment effect is greatest in the third and fifth decades of life, and is consistently patterned with microstructural and frequency differences previously reported by other researchers. It is suggested that δ¹⁵N is reflecting differences in urea excretion and the renal processing and clearance of calcium and phosphorus. The study not only alerts us to the susceptibility of stable nitrogen isotopes to non-dietary (i.e. physiological) factors, but also identifies nitrogen isotope ratios as a possible new marker for osteopenia. Am J Phys Anthropol 103:185–199, 1997. © 1997 Wiley-Liss, Inc.