Dark calcification and the daily rhythm of calcification in the scleractinian coral, <Emphasis Type="Italic">Galaxea fascicularis</Emphasis>

The rate of calcification in the scleractinian coral Galaxea fascicularis was followed during the daytime using ⁴⁵Ca tracer. The coral began the day with a low calcification rate, which increased over time to a maximum in the afternoon. Since the experiments were carried out under a fixed light intensity, these results suggest that an intrinsic rhythm exists in the coral such that the calcification rate is regulated during the daytime. When corals were incubated for an extended period in the dark, the calcification rate was constant for the first 4 h of incubation and then declined, until after one day of dark incubation, calcification ceased, possibly as a result of the depletion of coral energy reserves. The addition of glucose and Artemia reduced the dark calcification rate for the short duration of the experiment, indicating an expenditure of oxygen in respiration. Artificial hypoxia reduced the rate of dark calcification to about 25% compared to aerated coral samples. It is suggested that G. fascicularis obtains its oxygen needs from the surrounding seawater during the nighttime, whereas during the day time the coral exports oxygen to the seawater.     

Variation in calcification rate of Acropora downingi relative to seasonal changes in environmental conditions in the northeastern Persian Gulf

There is a strong interest in understanding how coral calcification varies with changing environmental conditions, especially given the projected changes in temperature and aragonite saturation due to climate change. This study explores in situ variation in calcification rates of Acropora downingi in the northeastern Persian Gulf relative to seasonal changes in temperature, irradiance and aragonite saturation state (Ω _arag). Calcification rates of A. downingi were highest in the spring and lowest in the winter, and intra-annual variation in calcification rate was significantly related to temperature (r ² = 0.30) and irradiance (r ² = 0.36), but not Ω _arag (r ² = 0.02). Seasonal differences in temperature are obviously confounded by differences in other environmental conditions and vice versa. Therefore, we used published relationships from experimental studies to establish which environmental parameter(s) (temperature, irradiance, and/or Ω _arag) placed greatest constraints on calcification rate (relative to the maximum spring rate) in each season. Variation in calcification rates was largely attributable to seasonal changes in irradiance and temperature (possibly ~57.4 and 39.7% respectively). Therefore, we predict that ocean warming may lead to increased rates of calcification during winter, but decelerate calcification during spring, fall and especially summer, resulting in net deceleration of calcification for A. downingi in the Persian Gulf.

     

Development of Vitellogenin-ELISA,an In Vivo Bioassay,and Identification of Two Vitellogenesis-Inhibiting Hormones of the Tiger Shrimp <Emphasis Type="BoldItalic">Penaeus monodon</Emphasis>

A sandwich enzyme-linked immunosorbent assay (ELISA) for the quantification of vitellogenin in the hemolymph of Penaeus monodon is reported. Lipovitellin from the mature ovary was purified using hydroxylapatite column chromatography and used as the standard protein, which was serially diluted from 3336 to 6.51 μg and gave a linear plot. Sensitivity results showed ELISA was insensitive to samples that did not contain vitellogenin or lipovitellin. Specificity results showed the degree of the discrimination of the assay between positive samples (having vitellogenin or lipovitellin) and negative samples (devoid of vitellogenin or lipovitellin). Reproducibility studies showed that the intraassay coefficient of variation was 5.1% (n= 11) and the interassay coefficient of variation was 5.15% (n= 13). Separation of X-organ sinus gland peptides using a reversed phase column gave a total of 37 fractions. Two fractions were found to reduce the hemolymph vitellogenin concentration in a time-dependent manner and could be identified as vitellogenesis-inhibiting hormones I and II. Received February 2, 2001; accepted June 24, 2001     

Interpopulation variation in fluctuating asymmetry of the palmar A-B ridge-count

Fluctuating asymmetry was measured by the Pearson correlation coefficient between a-b ridge-counts of the right and left hands. Such data were available for 38 samples, which were grouped by major geographical regions. Significant heterogeneity exists within some of the geographical groups but the between geographical group component is highly significant in both sexes. Our results show that a sizable fraction of variation in fluctuating asymmetry is related to geographical race. There is no evidence that it follows environmental or ecological lines. The variation between geographical races suggests that there is a genetic component in the magnitude of fluctuating asymmetry.     

Genetic diversity in pollen beetles (Meligethes aeneus) in Sweden: role of spatial,temporal and insecticide resistance factors

1 Pollen beetles Meligethes aeneus are pests of oilseed Brassica crops that are subject to intensive chemical control. Resistance to pyrethroids has been reported. Although this insect is of great economic importance, little is known about its genetic properties and population structure. 2 Amplified fragment length polymorphism (AFLP) analysis with the restriction endonuclease combination EcoRI and PstI was performed on 133 samples of groups of three pollen beetles collected during 2001–04 from five different provinces of Sweden. Both susceptible and resistant insects were studied. Using one primer combination, more than 450 polymorphic DNA fragments were obtained and, in total, four primer combinations were used for analysis. A subsample of 59 single beetles was analysed using one primer combination. 3 AFLP profiles were analysed by similarity measures using the Nei and Li coefficient and Neighbour‐joining dendrograms were generated. The dendrogram built using 133 samples showed three distinct groups, each containing beetles representing one generation. Statistical analysis using analysis of molecular variance of single beetle samples showed no evidence of significant genetic difference between resistant and susceptible beetles. Instead, a clear difference between samples, depending on time of collection and generation, was observed. 4 The expected regional population structure, although statistically significant, explained little of the variation. The levels of genetic variation within populations were very high. There appears to be a high rate of gene flow between pollen beetle populations. The implications of this in the context of insecticide resistance are discussed.     

Size and shape variation in the proximal femur of Australopithecus africanus

Aside from use as estimates of body mass dimorphism and fore to hind limb joint size comparisons, postcranial elements have not often contributed to assessments of variation in Australopithecus africanus. Meanwhile, cranial, facial, and dental size variation is interpreted to be high or moderately high. Further, the cranial base and face express patterns of structural (shape) variation, which are interpreted by some as evidence for the presence of multiple species. Here, the proximal femur is used to consider postcranial size and shape variation in A. africanus. Original fossils from Makapansgat and Sterkfontein, and samples from Homo, Pan, Gorilla, and Pongo were measured. Size variation was assessed by comparing the A. africanus coefficient of variation to bootstrapped distributions of coefficient of variation samples for each taxon. Shape variation was assessed from isometrically adjusted shape variables. First, the A. africanus standard deviation of log transformed shape variables was compared to bootstrapped distributions of logged standard deviations in each taxon. Second, shape variable based Euclidean distances between fossil pairs were compared to pairwise Euclidean distance distributions in each reference taxon. The degree of size variation in the A. africanus proximal femur is consistent with that of a single species, and is most comparable to Homo and Pan, lower than A. afarensis, and lower than some estimates of cranial and dental variation. Some, but not all, shape variables show more variation in A. africanus than in extant taxa. The degree of shape difference between some fossils exceeds the majority of pairwise differences in the reference taxa. Proximal femoral shape, but not size, variation is consistent with high estimates of A. africanus cranial variation.     

Linking genotype to phenotype in a changing ocean: inferring the genomic architecture of a blue mussel stress response with genome‐wide association

A key component to understanding the evolutionary response to a changing climate is linking underlying genetic variation to phenotypic variation in stress response. Here, we use a genome‐wide association approach (GWAS) to understand the genetic architecture of calcification rates under simulated climate stress. We take advantage of the genomic gradient across the blue mussel hybrid zone (Mytilus edulis and Mytilus trossulus) in the Gulf of Maine (GOM) to link genetic variation with variance in calcification rates in response to simulated climate change. Falling calcium carbonate saturation states are predicted to negatively impact many marine organisms that build calcium carbonate shells – like blue mussels. We sampled wild mussels and measured net calcification phenotypes after exposing mussels to a ‘climate change’ common garden, where we raised temperature by 3°C, decreased pH by 0.2 units and limited food supply by filtering out planktonic particles >5 μm, compared to ambient GOM conditions in the summer. This climate change exposure greatly increased phenotypic variation in net calcification rates compared to ambient conditions. We then used regression models to link the phenotypic variation with over 170 000 single nucleotide polymorphism loci (SNPs) generated by genotype by sequencing to identify genomic locations associated with calcification phenotype, and estimate heritability and architecture of the trait. We identified at least one of potentially 2–10 genomic regions responsible for 30% of the phenotypic variation in calcification rates that are potential targets of natural selection by climate change. Our simulations suggest a power of 13.7% with our study's average effective sample size of 118 individuals and rare alleles, but a power of >90% when effective sample size is 900.     

The mutual interplay between calcification and coccolithovirus infection

Two prominent characteristics of marine coccolithophores are their secretion of coccoliths and their susceptibility to infection by coccolithoviruses (EhVs), both of which display variation among cells in culture and in natural populations. We examined the impact of calcification on infection by challenging a variety of Emiliania huxleyi strains at different calcification states with EhVs of different virulence. Reduced cellular calcification was associated with increased infection and EhV production, even though calcified cells and associated coccoliths had significantly higher adsorption coefficients than non-calcified (naked) cells. Sialic acid glycosphingolipids, molecules thought to mediate EhV infection, were generally more abundant in calcified cells and enriched in purified, sorted coccoliths, suggesting a biochemical link between calcification and adsorption rates. In turn, viable EhVs impacted cellular calcification absent of lysis by inducing dramatic shifts in optical side scatter signals and a massive release of detached coccoliths in a subpopulation of cells, which could be triggered by resuspension of healthy, calcified host cells in an EhV-free, ‘induced media’. Our findings show that calcification is a key component of the E. huxleyi-EhV arms race and an aspect that is critical both to the modelling of these host–virus interactions in the ocean and interpreting their impact on the global carbon cycle.     

The importance of monitoring metabolic recovery in the coral Acropora cervicornis after short-term exposure to drilling muds: Calcification rate and protein concentration

The effect of used drilling muds on coral health was examined by monitoring changes in calcification rate and soluble tissue protein concentration in the coral Acropora cervicornis. Exposure to 25 ppm (v/v) of one mud for 24 h reduced calcification rate in the growing tips by as much as 62%. In recovery experiments, corals were exposed to drilling muds for 24 h; some of them were allowed to recover in clean seawater for 48 h. After the 24-hour exposure, calcification rates were significantly less than those of the controls. After a 48-hour recovery period, calcification rates returned to control levels for one mud but were still significantly below control levels for another. The results indicate that the capacity for recovery after exposure cannot be predicted from the results of experiments on exposure only. Recovery capacity must be independently verified for all studies on the effects of short-term exposure to drilling muds.     

Calcification rate and the effect of temperature in a zooxanthellate and an azooxanthellate scleractinian reef coral

Calcification rates, normalized to skeletal mass, in the zooxanthellate Galaxea fascicularis and the azooxanthellate Dendrophyllia sp. were similar over the whole temperature range of 18–29 °C. Calcification was measured by Ca⁴⁵ incorporation in corals that were naturally acclimated to the prevailing seawater temperature. In both species maximum calcification rate occurred at about 25 °C and calcification rates can be fitted to a Gaussian distribution with respect to temperature. The similarity in temperature dependence of the zooxanthellate and azooxanthellate coral suggests that temperature affects some fundamental process of calcification that is independent of light effects. It is shown that two different populations of Galaxea fascicularis have distinctly different ratios of tissue protein to skeletal mass per polyp. This indicates that tissue protein may not be suitable for normalizing calcification rates in individual coral polyps, both within and between species. Intra- and interspecific comparisons of calcification rates may be better made on the basis of skeletal mass when polyps are similar in size and shape.Communicated by Topic Editor C. Barnes     

Variation in synonymous substitution rates among mammalian genes and the correlation between synonymous and nonsynonymous divergences

Using mammalian gene sequences, the variances in the numbers of synonymous and nonsynonymous substitutions among genes were estimated together with the correlation coefficient between the two. The expected correlation coefficient can be obtained under the neutral theory using these estimated values of the variances. The expected coefficient is found to often be one-half to two-thirds of the observed value. Possible causes for the disagreement were discussed, such as correlated selective constraints on the two types of substitutions and excess doublet mutations. The variance of mutation rate and that of selective constraint were also estimated. The results show that the coefficient of variation of the former is 0.2–0.3, whereas that of the latter is 0.7–0.9. Correspondence to: T. Ohta     

Crystallization and Reconstitution of Yeast Aconitase

The combination of flow injection analysis and the electrochemical method was investigated for measuring the organic acid content of citrus fruits. The controlled-potential four-electrode method was used for conductance measurements. The instrumentation of the measuring apparatus used in this study is defined. The optimum conditions of the flow system were as follows: injection volume 35 μ1, mixing coil length 100 cm, flow rate 20ml/min and dilution ratio of samples 1:130. A linear correlation between the peak conductivity and the organic acid content was obtained with high correlation coefficient (r=0.992). It is possible to analyze about 50 samples within an hour using the method, and the coefficient of variation with any given juice for 20 assays was less than 0.5%.     

Dietary variation within and between populations of northeast Atlantic killer whales,Orcinus orca,inferred from δ13C and δ15N analyses

Epidermal skin samples from eastern North Atlantic killer whales, Orcinus orca, were analyzed for carbon and nitrogen stable isotope ratios. From those, comparisons within a data set of 17 samples collected from Tysfjord, Norway, in November suggested that diet is relatively specialized during this time period at this location. There were significant differences between a small set of samples from Iceland and those collected from Norway, which had all been assigned to the same population by a previous population genetics study. The results would be consistent with matrilines feeding on either the Norwegian or Icelandic stocks of Atlantic herring (Clupea harengus). There was no significant difference within Icelandic samples between those assigned to the population known to feed upon herring and those assigned to a population hypothesized to follow Atlantic mackerel (Scomber scombrus). The greatest differences were between the epidermal samples analyzed in this study and tooth and bone collagen samples from the North Sea that were analyzed previously, which also showed significantly more variation in isotopic ratios than found for skin samples. These differences could reflect differences in turnover rate, differences in diet‐tissue fractionation and discrimination due to the amino acid composition of the different tissues, and/or greater competition promoting dietary variation between groups in the North Sea.     

The transmissibility of a potyvirus by an aphid vector: reliability of measurements under controlled conditions

Reproducibility of measurements of the transmission rate of a given potyvirus by a given aphid vector was assessed using a standardised transmission procedure. The plant-virus-vector system studied was: Cucurbita pepo - Papaya ringspot virus - Aphis gossypii. Several elements concerning the rearing and handling of the aphids were analysed. One of these, the climatic conditions under which the aphids were reared, influenced the transmission rate of the virus. Six trials were carried out to measure the rate of transmission of the virus from a defined leaf surface area. The measurements appeared to be reliable. A two year experiment was carried out to compare transmission rates at different dates. Although strong variations were observed in some source plants, the transmission rates obtained for samples of 10 source plants were stable, with a 10% coefficient of variation. The results show that transmissibility of the virus is a stable property which is independent of the trial date. They are discussed in relation to standardised elements of the transmission process.     

Inter-simple sequence repeat (ISSR) variation in forest coffee trees (Coffea arabica L.) populations from Ethiopia

Genetic variation of forest coffee trees (Coffea arabica L.) from four regions of Ethiopia was investigated using inter-simple sequence repeat (ISSR) markers. A total of 160 individuals representing 16 populations were sampled. Eleven ISSR primers amplified a total of 123 fragments of which 31 fragments (25%) were polymorphic. Estimate of total gene diversity (H _T), and the coefficient of genetic differentiation (G _ST) were 0.37 and 0.81, respectively. This indicates that most of the variability is between populations than within populations. The partitioning of genetic variation into within and between populations based on Shannon’s information index also revealed more differentiation between populations (0.80) than within populations (0.20). In the phenogram most of the coffee tree samples were clustered on the basis of their regions of origin but failed to cluster according to their respective populations, which could be attributed to the presence of substantial gene flow between adjacent populations in each region assisted by man in the process of transplantation or by wild animals such as monkeys, which eat the berries and defecate the seeds elsewhere. On the other hand, the inter-regional clustering of some coffee tree samples from Bale and Jimma regions could be due to the transport of coffee seeds across regions and their subsequent planting. Although ISSR markers detected lower polymorphic loci than previously reported results with random amplified polymorphic DNA (RAPD) markers on the same materials, it can be used as an alternative method for molecular characterization of C. arabica populations. The results may provide information to select sites for in situ conservation.     

High performance liquid chromatographic determination of glycoalkaloids in callus and fruits ofSolanum eleagnifolium

Summary A method for the determination by HPLC of the triglycosides, solamargine and solasonine, in fruits and callus was developed. It was used to determine the content of triglycosides in samples fromSolanum eleagnifolium. The method involves ion-pair extraction of glycoalkaloids, clean-up of the samples and HPLC analysis. Recoveries ranged between 94–98%, and the coefficient of variation was approximately 5%.     

Seasonal Variation in Shell Calcification of Planktonic Foraminifera in the NE Atlantic Reveals Species-Specific Response to Temperature,Productivity, and Optimum Growth Conditions

Using shells collected from a sediment trap series in the Madeira Basin, we investigate the effects of seasonal variation of temperature, productivity, and optimum growth conditions on calcification in three species of planktonic Foraminifera. The series covers an entire seasonal cycle and reflects conditions at the edge of the distribution of the studied species, manifesting more suitable growth conditions during different parts of the year. The seasonal variation in seawater carbonate saturation at the studied site is negligible compared to other oceanic regions, allowing us to assess the effect of parameters other than carbonate saturation. Shell calcification is quantified using weight and size of individual shells. The size–weight scaling within each species is robust against changes in environmental parameters, but differs among species. An analysis of the variation in calcification intensity (size-normalized weight) reveals species-specific response patterns. In Globigerinoides ruber (white) and Globigerinoides elongatus, calcification intensity is correlated with temperature (positive) and productivity (negative), whilst in Globigerina bulloides no environmental forcing is observed. The size–weight scaling, calcification intensity, and response of calcification intensity to environmental change differed between G. ruber (white) and G. elongatus, implying that patterns extracted from pooled analyses of these species may reflect their changing proportions in the samples. Using shell flux as a measure of optimum growth conditions, we observe significant positive correlation with calcification intensity in G. elongatus, but negative correlation in G. bulloides. The lack of a consistent response of calcification intensity to optimum growth conditions is mirrored by the results of shell size analyses. We conclude that calcification intensity in planktonic Foraminifera is affected by factors other than carbonate saturation. These factors include temperature, productivity, and optimum growth conditions, but the strength and sign of the relationships differ among species, potentially complicating interpretations of calcification data from the fossil record.     

Early ontogeny of Adinia xenica (Pisces,Cyprinodontiformes): 2. Implications of embryonic resting interval for larval development

Synopsis Larval development commences with first exogenous feeding, and ends with final remodelling of caenogenetic structures into the definitive organs of juvenile and adult. For the intertidal cyprinodontid Adinia xenica this generally corresponds to the interval between hatching and completion of scalation. The final step of the embryo period is a resting interval of variable duration. Embryos were induced to hatch after 2 and 10 days of this near arrest. Although the general pattern of larval development was the same for both groups, differences were observed in the rates and order of calcification of skeletal elements, fin differentiation and growth, and scalation. For example, embryos hatching 8 days later in the resting interval were already pattially calcified, but completed calcification at a slower rate than the group hatching after 2 days. These differences may be due to effects of the duration of the resting interval itself; or they may reflect genetic variation of which age at hatching is only one manifestation.     

A coral polyp model of photosynthesis, respiration and calcification incorporating a transcellular ion transport mechanism

A numerical simulation model of coral polyp photosynthesis, respiration and calcification was developed. The model is constructed with three components (ambient seawater, coelenteron and calcifying fluid), and incorporates photosynthesis, respiration and calcification processes with transcellular ion transport by Ca-ATPase activity and passive transmembrane CO₂ transport and diffusion. The model calculates dissolved inorganic carbon and total alkalinity in the ambient seawater, coelenteron and calcifying fluid, dissolved oxygen (DO) in the seawater and coelenteron and stored organic carbon (CH₂O). To reconstruct the drastic variation between light and dark respiration, respiration rate dependency on DO in the coelenteron is incorporated. The calcification rate depends on the aragonite saturation state in the calcifying fluid (Ωa _cal). Our simulation result was a good approximation of “light-enhanced calcification.” In our model, the mechanism is expressed as follows: (1) DO in the coelenteron is increased by photosynthesis, (2) respiration is stimulated by increased DO in the light (or respiration is limited by DO depletion in the dark), then (3) calcification increases due to Ca-ATPase, which is driven by the energy generated by respiration. The model simulation results were effective in reproducing the basic responses of the internal CO₂ system and DO. The daily calcification rate, the gross photosynthetic rate and the respiration rate under a high-flow condition increased compared to those under the zero-flow condition, but the net photosynthetic rate decreased. The calculated calcification rate responses to variations in the ambient aragonite saturation state (Ωa _amb) were nonlinear, and the responses agreed with experimental results of previous studies. Our model predicted that in response to ocean acidification (1) coral calcification will decrease, but will remain at a higher value until Ωa _amb decreases to 1, by maintaining a higher Ωa _cal due to the transcellular ion transport mechanism and (2) the net photosynthetic rate will increase.     

Comparative application of the coefficient of variation and range-based statistics for assessing the taxonomic composition of fossil samples

Dental variation has been used commonly to assess taxonomic composition in morphologically homogeneous fossil samples. While the coefficient of variation (CV) has been used traditionally, range-based measures of variation, such as the range as a percentage of the mean (R%) and the maximum/minimum index (I_max/min) have recently become popular alternatives. The current study compares the performance of these statistics when applied to single- and pooled-species dental samples of extant Cercopithecus species. A common methodology for such problems of species discrimination has been to simply compare the maximum value of a variation statistic observed in extant samples with that observed in the fossil sample. However, regardless of what statistic is used, this approach has an unknowable Type I error rate, and usually has low power to detect multiple species. A more appropriate method involves a formal hypothesis test. The null hypothesis is that the level of variation in the fossil sample does not exceed what might be expected in a sample drawn randomly from a reference population, taking into account sampling error and the size of the fossil sample. Previous research using this method with the CV has indicated that it offers considerable power at an acceptable Type I error rate. In the current study, the data of primary interest were posterior dental dimensions for single- and pooled species samples from extant Cercopithecus species. In addition, the study also investigated the relative performance of variation statistics when applied to highly dimorphic canine dimensions, since much recent work has employed sexually dimorphic dental dimensions for assessing single-species hypotheses. The results indicate that the CV consistently out-performed the range-based statistics when using posterior dental dimensions to test a single-species hypothesis. Regardless of which statistic was used, tests on sexually dimorphic dimensions offered minimal power. In consideration of these results and the problem of studywise Type I error rates, we recommend against the use of multiple measures of variation to test for multiple species composition, and advocate the CV as the statistic of choice when using the method of Cope & Lacy (1992). For similar reasons, we argue for careful selection of dental variables for inclusion in such analyses, and in particular recommend against including sexually dimorphic dimensions when testing for multiple species composition.