Cardenolide content,emetic potency,and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus,and their larval host-plant milkweed,Asclepias humistrata,in Florida

Summary This paper is the fourth in a series on cardenolide fingerprints of monarch butterflies (Danaus plexippus, Danainae) and their host-plant milkweeds (Asclepiadaceae) in the eastern United States. Cardenolide concentrations ofAsclepias humistrata plants from north central Florida ranged from 71 to 710 µg/0.1 g dry weight, with a mean of 417 µg/0.1 g. Monarchs reared individually on these plants contained cardenolide concentrations ranging from 243 to 575 µg/0.1 g dry weight, with a mean of 385 µg/0.1 g. Cardenolide uptake by butterflies was independent of plant concentration, suggesting that sequestration saturation occurs in monarchs fed cardenolide-rich host plants. Thinlayer chromatography resolved 19 cardenolides in the plants and 15 in the butterflies. In addition to humistratin,A. humistrata plants contained several relatively non-polar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. These produced a butterfly cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. In emetic assays with the blue jay,Cyanocitta cristata, the 50% emetic dose (ED₅₀) per jay was 57.1 µg, and the average number of ED₅₀ units per butterfly was 13.8, establishing that this important south eastern milkweed produces highly emetic, chemically defended monarchs. Our data provide further support for the use of cardenolide fingerprints of wild-caught monarchs to make ecological predictions concerning defence against natural enemies, seasonal movement and larval host-plant utilization by monarch butterflies during their annual cycle of migration, breeding and overwintering.     

Filamentous cyanophytes containing phycourobilin and in symbiosis with sponges and an ascidian of coral reefs

A study was made of the ultrastructure and pigment composition of filamentous cyanophytes living in symbiosis with several sponges and a colonial didemnid ascidian collected from the southern end of the Great Barrier Reef, Australia, between 1983 and 1986. The sponges were Dysidea herbacea Keller and several other encrusting sponges which have not been identified; the ascidian was Trididemnum miniatum Kott (1977). The cyanophyte Oscillatoria spongeliae (Shultz) Hauck was identified as the symbiont of several of the sponges, including D. herbacea. Two other unidentified Oscillatoria species were found in a bristly papillate sponge and in T. miniatum. Chlorophyll a, alone, was present in all the symbionts with the exception of T. miniatum, which contained the cosymbiont Prochloron and where chlorophyll b was also present. Two phycoerythrins were isolated by chromatography and chromatofocusing. Both resembled C-phycoerythrin, but one of the two carried the chromophore phycourobilin as well as phycoerythrobilin possibly on both the and subunits, which had apparent molecular masses of 18 and 20 kdaltons. No subunit was present. Ultrastructurally, the three Oscillatoria species were distinguished by an unusual type of parallel, longitudinal, thylakoid organisation; the arrangement was different in detail in each species.     

Microbial reaction rates and bacterial communities in sediment surrounding burrows of two nereidid polychaetes (Nereis diversicolor and N. virens)

The effects of infaunal mode of life on sediment properties, microbial reaction rates, as well as abundance and composition of bacterial communities were studied in sediment surrounding burrows (mucus lining, oxidised wall, ambient anoxic and surface sediment) of two closely related, but behaviourally different, nereidid polychaete worms: the facultative suspension-feeder Nereis (Hediste) diversicolor and the obligate deposit-feeder Nereis (Neanthes) virens. Burrow sediment of the two species was collected from two adjacent (50 m distance) shallow sandy locations (Kertinge Nor, Denmark). The burrow lining and wall of both polychaete species were enriched in organic matter originating from mucous secretions by the inhabitants and phytoplankton trapped through irrigation. This was more evident for N. diversicolor that shows a significantly higher irrigation rate than N. virens. Both the organic matter mineralisation rates (based on anaerobic incubations) and bacterial abundance were higher along the burrow linings and walls. However, accumulation of porewater TCO₂ and dissolved organic carbon in sediments adjacent to burrows increased most rapidly in the presence of N. diversicolor, suggesting higher heterotrophic activity associated with this species. Surprisingly, bacterial abundance was lower around burrows of N. diversicolor than those from N. virens indicating that burrow environments from the first species harbour a more active bacterial community. Molecular fingerprints of the 16S rRNA gene from bacterial communities showed that the composition of the burrow linings and walls resembled the ambient anoxic sediment rather than the oxic sediment surface. On the other hand, the bacterial fingerprints of the sediment surrounding the burrows of the two polychaete species were markedly different suggesting either a site-specific difference in sediment parameters or a significant species-specific impact of the burrow inhabitants.     

Primary production as influenced by diel periodicity of phytoplankton photosynthesis

Diel periodicity in parameters of photosynthesis-irradiance (P-I) curves was incorporated into calculations of integral daily phytoplankton production for the Santa Barbara Channel off southern California (USA). Model equations of the relationship between photosynthesis and light were used in combination with observed slope () and asymptote (P _max) values presented in the preceding paper. Primary production was always 19 to 39% less than comparable estimates obtained with the assumption of constant maximum daily and P _max values. Regardless of which P-I formulation was used of 6 tested, observed production (using a temporal series of simulated in situ incubations) ranged from 13% less to 25% more than estimates from constant midday and P _max values. The amplitude and timing of diel oscillations differed somewhat among 3 field stations. Maximum to minimum ratios ranged from approximately 3 to 5 for , and 4 to 6 for P _max. The differences in amplitude and timing of oscillations in P-I curves both contributed to errors in calculating phytoplankton production. Thus, photosynthetic periodicity in the upwelling area of the Santa Barbara Channel influences phytoplankton production. There were oscillations in both and P _max, and the time-dependence of these parameters should be considered to improve the accuracy of predictive models of primary productivity.     

Characteristics of dispersions and water-soluble extracts of crude and refined oils and their toxicity to estuarine crustaceans and fish

The quantitative hydrocarbon composition and behavior in seawater of water-soluble fractions (WSF) and oil-in-water dispersions (OWD) of 4 oils was investigated. Two crude oils, South Louisiana crude and Kuwait crude, and two refined oils, No. 2 fuel oil and bunker C residual oil, were used in these investigations. The WSFs of the crude oils had higher total oil-hydrocarbon concentrations and were richer in light aliphatics and single-ring aromatics than were the WSFs of the refined oils. The WSFs of the refined oils contained significantly higher concentrations of naphthalenes than did those of the crudes. The hydrocarbon composition of the aqueous phase of OWDs closely resembled that of the parent oils. Gentle aeration of the OWDs resulted in a loss of 80 to 90% of the aqueous hydrocarbons in 24 h. Alkanes disappeared from the dispersions more rapidly than aromatics. The WSFs and OWDs of the refined oils were considerably more toxic to the 6 test species than were those of the crude oils. The test species can be ranked according to increasing sensitivity to oil as follows: Cyprinodon variegatus, Menidia beryllina, Fundulus similus, Penaeus aztecus postlarvae, Palaemonetes pugio and Mysidopsis almyra. The results of this investigation are discussed in relation to the potential impacts of oil spills on the marine and estuarine environments.     

Effects of Habitat Area on the Distribution of Grassland Birds in Maine

We used multiple and logistic regression analysis to study the breeding-area requirements of 10 species of grassland and early-successional birds at 90 grassland-barren sites in Maine. The incidence of six of the species was clearly sensitive to the area of grassland. Upland Sandpipers, the species with the largest area requirements, were infrequent at sites of less than 50 ha and reached 50% incidence at those of about 200 ha. Grasshopper Sparrows reached 50% incidence at about 100 ha, Vesper Sparrows at about 20 ha, and Savannah Sparrows at about 10 ha. Incidence for three edge species, Brown Thrasher, Common Yellowthroat, and Song Sparrow, was negatively correlated with open area, and incidence for Field Sparrows was not strongly influenced by grassland size. These results indicate that grasslands need to be approximately 200 ha in area if they are to be likely to support a diverse grass land bird fauna. However, large grasslands or grassland-barrens are rare; random samples indicated that in Maine only 1% of hayfields and only 8% of grassland-barrens were more than 64 ha in area. Conservation efforts seeking to protect habitat for rare grassland birds need to consider sites of at least 100 ha—and preferably 200 ha—in size, and these are notably rare in Maine and probably throughout New England and eastern North America. Airports provide some of the last extensive patches of grassland habitat in the northeast. To maintain viable populations of area-sensitive grassland birds, management of these sites for nesting birds will become increasingly important.     

Compartmentation and turnover of organic carbon in the Staghorn coral Acropora formosa

Four colonies of Acropora formosa were incubate with Na₂ ¹⁴CO₃ for separate 2 h periods within a 24 h period, and then returned to the reef from which they were collected. Terminal branches were collected at intervals over the following 5 d and analysed for radioactivity associated with the skeleton and certain organic pools. Colonies incubated at night showed little or no loss of fixed radioactivity during the 5 d on the reef. However, 50–60% of photosynthetically-fixed ¹⁴C was lost from the terminal branches during the first 40 h on the reef. This loss of radioactivity probably resulted from release of mucus and dissolved organic carbon from the coral tissues. Most of the loss of photosynthetically-fixed ¹⁴C was due to decrease in the radioactivity of lipids (80% of the total ¹⁴C loss) and methanol-water soluble compounds. Determination of any sequencing in metabolic compartments was made difficult by the rapidity with which ¹⁴C dissappeared from most of the metabolic pools measured. ¹⁴C was incorporated into the skeleton throughout the 5 d on the reef, although the rate of incorporation was very low in colonies which had been incubated with Na₂ ¹⁴CO₃ at night.     

Measuring concentrations of volatile organic compounds in vinyl flooring

The initial solid-phase concentration of volatile organic compounds (VOCs) is a key parameter influencing the emission characteristics of many indoor materials. Solid-phase measurements are typically made using solvent extraction or thermal headspace analysis. The high temperatures and chemical solvents associated with these methods can modify the physical structure of polymeric materials and, consequently, affect mass transfer characteristics. To measure solid-phase concentrations under conditions resembling those in which the material would be installed in an indoor environment, a new technique was developed for measuring VOC concentrations in vinyl flooring (VF) and similar materials. A 0.09-m2 section of new VF was punched randomly to produce -200 0.78-cm2 disks. The disks were milled to a powder at -140 degrees C to simultaneously homogenize the material and reduce the diffusion path length without loss of VOCs. VOCs were extracted from the VF particles at room temperature by fluidized-bed desorption (FBD) and by direct thermal desorption (DTD) at elevated temperatures. The VOCs in the extraction gas from FBD and DTD were collected on sorbent tubes and analyzed by gas chromatography/mass spectrometry (GC/MS). Seven VOCs emitted by VF were quantified. Concentration measurements by FBD ranged from 5.1 microg/g VF for n-hexadecane to 130 microg/g VF for phenol. Concentrations measured by DTD were higher than concentrations measured by FBD. Differences between FBD and DTD results may be explained using free-volume and dual-mobility sorption theory, but further research is necessary to more completely characterize the complex nature of a diffusant in a polymer matrix.     

Feeding preferences and the relationships between food choice and assimilation efficiency in the herbivorous marine snail Lithopoma undosum (Turbinidae)

Preference rankings for 13 macrophytes were established for the subtidal herbivorous snail Lithopoma undosum using two-choice laboratory experiments and consumption rates. L. undosum did not discriminate among three kelp foods (Egregia menziesii, Eisenia arborea and Macrocystis pyrifera) but ate kelp preferentially and more rapidly over all but Ulva spp. among tested macrophytes. Secondary preferences were established for the red alga Pterocladiella capillacea, followed by the coralline Lithothrix aspergillum, whereas the brown seaweeds Zonaria farlowii and Halidrys dioica and the seagrass Phyllospadix torreyi were the least preferred macrophytes. Fastest consumption rates (1.91 g day⁻¹) were measured in trials consisting only of kelp foods. These results indicate that L. undosum exhibits clear feeding preferences even when given less-preferred, non-kelp macrophytes. Using an ash-marker technique, we determined total organic, carbon, and nitrogen assimilation efficiencies (AE%) for six macroalgae used in preference trials. Tested macrophytes were assimilated at different efficiencies but a pattern was not detected between AE (%) and a macrophyte’s position in L. undosum’s preference hierarchy. Highest total organic AEs were found for P. capillacea (61.2%) and H. dioica (59.4%); lowest AEs were detected for E. menziesii (34.9%), a preferred dietary item. Nitrogen was assimilated from red algae with higher efficiencies (74.9–84.3%) than from brown or green algae. These data suggest that the digestive capabilities of L. undosum are better suited for assimilating organic material and nitrogen from less-preferred, non-kelp foods. This supports the hypothesis that factors besides nutritional composition and digestive optimization have played a role in the evolution of feeding preferences in L. undosum and probably other herbivorous snails associated with northeastern Pacific kelp beds.