Seasonal production of moulting hormone in the barnacle Semibalanus balanoides

An analytical procedure for the quantification of ecdysteroids (crustacean moulting hormone) in barnacles was devised so that minimum sample size could be used. A combination of solvent partitions, Sephadex chromatography, silylation and gas chromatography with electron capture detection was devised, enabling ecdysteroids to be determined down to 20 pg. This was used to determine the amount of moulting hormone in a population of barnacles over a 30 month period. Levels varied from barely detectable in winter months to a maximum value of 1.5 μg kg^− 1 of wet weight of barnacles in September. Polar conjugates of 20-hydroxyecdysone were detected only during the winter months. The number of barnacles moulting at any time corresponded roughly to the titre of hormone present at that time.     

Regeneration of the penis in Balanus balanoides (L.)

In Balanus balanoides (L.) the penis is ‘lost’ each autumn. The regeneration of the penis is described. The primordium contains mesenchyme. The pedicel structures develop early as thickened areas of the exoskeleton and give rise to the characteristic structural features of the mature pedicel. Tissue differentiation begins in the pedicel and later extends to the penis proper which becomes annulated. The ductus eventually opens by a terminal invagination of the exoskeleton.     

Early changes in the shell form of Balanus balanoides (L.)

The shell shape of isolated growing individuals of Balanus balanoides (L.) changes considerably during early growth but by the time it reaches 3 mm in length its shape is almost the same as that of the adult, and thereafter it grows uniformly in all dimensions. The early metamorphosed barnacle is extremely squat with nearly vertical parietes and a relatively large aperture. The base, initially elliptical, becomes more circular and later develops ribbed borders. The carinolateral compartments are absent in the newly metamorphosed barnacle; their development and expansion is probably the main factor in the change in shape of the base. The apex of the carina, initially the lowest, becomes the highest point of the shell. The apex of the rostrum becomes relatively depressed during growth. The angle between the parietes and the base lies between 60° and 80° in newly metamorphosed individuals and drops to 35–45° in the adult.We suggest that the significance of these changes, if considered in conjunction with the orientation to current, is to maximize adhesion and minimize the drag of water currents on the shell, and to direct the water flow into the cirral net.     

Variation in penis morphology and mating ability in the acorn barnacle, Semibalanus balanoides

I examined variation in penis morphology of the acorn barnacle, Semibalanus balanoides, at different aggregation densities and at different levels of wave exposure. Barnacles in sparse, un-crowded aggregations had significantly longer penises than those from densely crowded groups, suggesting a response to increase the chance of reaching distant mating partners. Barnacles exposed to oceanic waves had penises with significantly greater basal diameter, possibly to strengthen the penis and retain function in turbulent conditions. I compared the percentage of individual barnacles with fertilized broods over a range of distances to their nearest possible mate in sites exposed to or protected from waves. As neighbor distance increased, the proportion of individuals with fertilized egg masses decreased in both wave-exposed and protected sites. However, at greater mate distances in the wave exposed sites, the proportion of individuals with fertilized eggs was significantly lower than the proportion in protected sites, indicating that exposure to waves hinders mating with neighbors at increasing distances. These results suggest that the intensity of mate competition may differ for barnacles between environments with different levels of wave exposure. These differences in male ability are predicted to alter relative sex allocation to male and female function.     

Wound-stimulated moulting and calcification responses in the adult cirripede Balanus balanoides (L.)

Earlier studies of hormonal action in adult barnacles have involved the injection of crustecdysone into the basal mantle space through a hole drilled in the shell. The drilling and injection result in increased moulting activity and in calcification within the mantle tissue around the site of injection. A preliminary investigation of the nature and causes of these responses has been carried out. The increase in moulting activity is distinct from that induced by injected crustecdysone and can be sustained over several months, the level of stimulation depending upon whether the adults are subjected to single or multiple injections. The associated calcification extends from the shell wall and develops as a layer around an area of wound tissue at the injection site. It appears that these moulting and calcification activities are healing responses to wounding and that the processes involved may be similar to those of other arthropods. Moulting in barnacles is apparently controlled by an ecdysone hormonal system and these results suggest that such a system may be implicated in the control of calcification.     

The sterols of Balanus balanoides (L.): The effect of controlled diets

The effect of a range of diets with varying cholesterol content on the sterol composition of the barnacle Balanus balanoides (L.) has been studied.The animals appear able to maintain their sterol composition for considerable periods (4 months), both quantitatively and qualitatively, in the absence of dietary sterols. This suggests either an extremely slow metabolic turnover or some capacity for de novo synthesis of sterols from other biochemical components.     

The fine structure and possible function of the sensory setae of the penis of Balanus balanoides (L.)

The structure and innervation of the sensory setae which are present in large numbers on the penis of Balanus balanoides (L.) have been established by scanning and transmission electron microscopy. Each seta contains a small number of sensory dendrites surrounded by an extracellular supporting tube which is presumed to be secreted by the enclosing sheath cell. The dendrites, which distally extend beyond both the sheath cell and supporting tube, terminate at the tip of the seta within a pore-like invagination of the cuticle and thus are in direct contact with the environment. Proximally bundles of dendrites pass into the penis tissue where they are surrounded by several sheath cells. The supporting tube terminates at a point within the body of the penis where a series of intracellular rods arise. The ciliary character of the dendrites is evident in this region, the microtubules being organized into the (9 × 2) +2 pattern. It is deduced that the sensilla are chemosensory; their structure is compared with that of other crustacean sensilla which are presumed to be chemosensory.     

Studies in the biochemistry of cirripede eggs. IV. The free Amino-acid pool in the eggs of Balanus balanoides (L.) and B. balanus (L.) during development

Changes in the ‘free’ amino acids, betaine, and trimethylamine oxide during the development of the eggs of Balanus balanoides (L.) and B. balanus (L.) have been determined; the results are given in terms of μM/g dry wt, μM/g water, and μM/10⁶ eggs. The amino acids are derived from the yolk proteins the net composition of which is known. Free amino acids are present in considerable quantity, as is commonly the case with crustacean tissue. Changes in the individual amino acids are discussed. B. balanus eggs contain large, and relatively constant, amounts of sarcosine; its function is unknown but large quantities are present in the more highly evolved cirripedes so far examined. A possible relation between betaine glycine, and sarcosine relative to choline metabolism is considered. Large amounts of taurine are present. There is a striking increase in β-alanine in the late stages of development; in B. balanoides it comes to be the most, and in B. balanus the third most common amino acid; its possible involvement in purine metabolism is considered. The relation between the amounts of the various entities in the eggs and in the bodies of the adult are examined.     

Seasonal changes in the activity of certain enzymes in crude tissue extracts of two common cirripedes, Balanus balanoides (L.) and B. balanus (L.).

The activity of glutamate-oxaloacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (ICDH), and aldolase in tissue extracts of Balanus balanoides (L.) and B. balanus (L.) have been determined at regular intervals over a year, and an attempt made to relate the results to known biochemical changes in the animals.The work was ecologically orientated. Preliminary studies on the effects of extractant, dialysis, and possible endogenous interference were made and the subsequent procedure rigorously standardized.Transamination was investigated by TLC. LDH and aldolase show striking seasonal changes; an abundance of the former is associated with the production of semen. MDH, concerned in carbohydrate and lipid metabolism, was present in considerable quantities but there were no marked seasonal changes. GPT activity is high when reproductive products are being produced from reserves of food and the activity is also related to the production of semen. GOT showed no clear seasonal trends.The results are discussed relative to the metabolism of crustaceans: the part played by proteins and lipids is stressed.     

Changes in the biochemical composition of Balanus balanoides (L.) under experimental conditions: The effect of a starch diet

A comparison has been made of the changes in biochemical composition of experimental and natural populations of Balanus balanoides (L.). The ‘bodies’ and the ovary were separately analysed. The lipid components have been investigated in some detail; lipid classes were separated and the component fatty acids of the major components (triglycerides + diglycerides and phospholipids) determined.Animals fed on brine shrimp increased in weight more than the natural population, that of the starved animals was less, and that of animals fed on starch only, even less. The starch-fed animals continued to produce faecal pellets and it is suggested that the greater energy expenditure in feeding (cirral) activity with little gain of useful metabolic substrate was responsible for this decrease in body weight. These differences extend to the component lipid classes. Even so, these starch-fed animals produced ovarian tissue as, indeed, did starved animals, presumably by transfer of body material; the low level or absence of nutrients did not interfere with the hormonal stimulation of reproduction.     

The early development of the cement apparatus in the barnacle, Balanus balanoides (L.) (Crustacea: Cirripedia)

The development of the cement cells of Balanus balanoides (L.) has been followed over a 26-day post-settlement period. Cells of the cyprid cement glands de-differentiate to form part of the juvenile barnacle apparatus, but the majority of juvenile cement cells develop by differentiation of collecting duct cells. These latter cement cells are recognized by apical invaginations which are continuous with the collecting duct lumen. Collecting ducts are present in the cyprid. The cement duct system was not studied in detail, but preliminary observations of the arrangement in B. balanoides and Elminius modestus Darwin have shown them to differ, the system in E. modestus being simpler.     

Influence of oil shale on intertidal organisms: Effect of oil shale extract on settlement of the barnacle Balanus balanoides (L.)

As part of a study to investigate the effect of oil seeps on intertidal organisms, oil extracts of Blackstone oil shale from Kimmeridge on the Dorset coast were used in laboratory experiments to test their effect on the settlement of the barnacle Balanus balanoides (L.). Thin films of oil extract painted on the surface of pits in slate panels had no effect on cyprid settlement when applied up to a surface density of 2.8 g · m^?2, representing a thickness of 3.3 μm. Larger surface densities of oil stimulated cyprids to settle in far greater numbers than on unoiled panels. The maximum effect was obtained at a surface density of between 14.0 and 56.0 g · m ^?2, representing a thickness of 16.5 μm and 66.0 μm. With higher concentration of oil in the pits, stimulation to settle was reduced although cyprid settlement was still encouraged at a surface density of oil of 112g · m^?2 or 132 μm thickness.The unfractionated crude oil shale extract was a less powerful stimulus for barnacle settlement than a partially purified solution of the integumental protein arthropodin, another strong settlement inducer for barnacle cyprids.     

The utilization of various biochemical entities in gonadally mature Balanus balanoides (L.) under starvaton and feeding in the absence of copulation

Copulation was prevented by isolation in Balanus balanoides (L.) and the changes in total body weight, carbohydrate, lipid, protein, RNA, and DNA followed when the animals were starved and fed; the results are compared with those on animals allowed to copulate with the consequent loss of semen and laying down of egg lamellae. General body tissues, semen, and ovaries are found not to be isolated compartments; on starvation both semen and ovary contribute to general metabolism, the former to a greater extent. Even when the isolated animals are fed there is still an almost complete resorption of semen, but the ovary is largely maintained. Possible control mechanisms are discussed.     

Tolerances of Nauplii of Balanus balanoides (L.) and Elminius modestus (Darwin) exposed to cyclical temperature fluctuations

1. 1.|Newly-released larvae of both E. modestus and B. balanoides show similar tolerances to steady temperature levels.

2. 2.|Both species show an ability to withstand a wider range of temperatures when subjected to cyclic thermal shocks.

3. 3.|E. modestus shows a greater tolerance to short-term thermal shocks than B. balanoides.

Dormancy of Medicago marina (L.) seed

Medicago marina (L.) is a Mediterranean species whose seeds show strong dormancy that prevents germination. We used an integrated approach of physiological analyses and proteomics to investigate the mechanisms that control M. marina dormancy/germination and that underlie stress tolerance. First, we evaluated the effects on dormancy breaking of the following treatments: mechanical scarification, freezing at −20 °C, storage for 4 months and heating at 100 °C for 1 h. Mechanical scarification and freezing were the most effective treatments in overcoming dormancy. The role of abscisic acid (ABA) in M. marina dormancy was studied by ELISA immuno-enzymatic assay. The ABA content of germinated and non-germinated mature (control) and treated seeds was determined. The level of ABA was higher in treated seeds than in control seeds; the most significant increase occurred in the heated seeds. A comparison of the ABA level in the germinated, control and treated seeds suggests that different mechanisms modulate ABA content in response to different stresses, and that a specific ABA-signalling pathway regulates germination. Proteomic analysis revealed 46 proteins differentially expressed between treated and untreated seeds; 14 of these proteins were subsequently identified by mass spectrometry. Several of the proteins identified are important factors in the stress response, and are involved in such diverse functions as lipid metabolism, protein folding and chromatin protection. Lastly, an analysis of the phosphoproteome maps showed that the function of many proteins in seeds subjected to temperature treatment is modulated through post-translational modifications.     

The rate of development of the embryos of Balanus balanoides (L.) from a number of european and American populations and the designation of local races

A summary is given of the distribution of Balanus balanoides (L.) and of earlier work on differences — racial and otherwise. The variation in the date at which different populations are fertilized is considered. Data are given on the time required for the development of the embryos of this species in vitro at 10 °C, with eggs from a number of populations from the eastern and western Atlantic and examined soon after collection, and also after transplantation and maintenance under ambient laboratory conditions over the next breeding season. Both aspects of reproduction are under genetic control. There is no correlation between the date of fertilization and the time required for development.Oocyte and egg size appear to be largely controlled by temperature.On both sides of the Atlantic there is a general tendency for the time of development to decrease with decreasing latitude. The population at Millport, Scotland appears unusual. The southernmost populations on European shores resemble those at the extreme south in North America.A study of the effect of temperature has shown that at lower temperatures the time of development of the embryos in vitro from all populations tends to be the same; the difference lies in the Q₁₀ values.The results are discussed relative to palaeontological data and the existence of clines in other features.     

Hoarding by honeybees (Apis mellifera L.)

The food hoarding by groups of fifty bees kept in small cages and provided with sugar syrup was studied. Less food was stored in a new comb than in an old one, whether the old comb had been used for storing food or rearing brood, and there was less in drone than in worker combs. The presence of light, larvae and the odour of honey discouraged storage of syrup, but the presence of a queen encouraged it. The amount stored also varied with the environmental temperature, the age of the bees concerned and with their previous physiological and behavioural experience including food deprivation and length of confinement. Increased food in honeystomachs sometimes compensated for less stored in combs.     

Tenacity and shell shape in six Patella species: Adaptive features

Measurements were made of te forces necessary to detach various South African Patella spp. These ranged from 5.18 to 1.95 kg/cm², with significant differences between the species. P. cochlear Born. had the highest value, followed in sequence by P. argenvillei Kr., longicosta Lam., P. granularis L., P. granatina L., and P. oculus Born. Adhesion is the only mechanism capable of providing forces of this magnitude.Differences between the species are related to differences in their morphology, high tenacity being associated with low mucus secretion, small number of mucocytes, and inflexibility of the foot. This is associated with a large area of muscle attachment on the shell and the size of basal haemocoelic spaces.Features favouring high tenacity conflict with those favouring mobility, and the limpets fall into two groups. P. granularis, P. oculus, and P. granatina occur on the upper shore and the last two usually occur in areas which are sheltered from strong wave action. They a rapid growth and high gonadal output, demanding extensive foraging and hence mobility at the cost of tenacity. P. cochlear and P. longicosta are territorial and together with P. argenvillei, remain low on the shore, grow slowly, and have a low reproductive output. Feeding is localized and mobility sacrificed for high tenacity which is essential in P. cochlear and P. argenvillei as they occur in areas of strong wave action.Shell height is not correlated with tenacity nor with the intensity of wave action normally experienced by each species, but P. cochlear, P. argenvillei and to a lesser extent P. granularis, are subjected to strong wave action and have proportionally narrower shells (increasing streamlining) and low coefficients of drag. The latter are low due to the rough but regular texture of the shells creating a turbulent boundary layer and hence reducing drag.     

The development of the retinomotor reactions in larval plaice (Pleuronectes platessa, L.) and turbot (Scophthalmus maximus, L.)

The development of the retinomotor reaction was examined histologically in the larvae of plaice (Pleuronectes platessa, L.) and turbot (Scophthalmus maximus, L.). In plaice retinal pigment migration is present from hatching, but, in turbot, does not occur until just before metamorphosis. In both species rods first develop at metamorphosis. Plaice show cone migration, but this does not occur in turbot. During dark-adaptation retinal pigment contraction started at ≈ 100 lux in all stages of plaice. Before metamorphosis pigment was fully contractedimmediately below 1000 lux, but after metamorphosis full contraction did not occur until the light intensity had fallen to < 1.0 lux. The thresholds for cone migration in plaice showed similar changes, although the actual light intensities were about an order of magnitude lower. In turbot there was always an abrupt transition between light- and dark-adaptation; the thresholds for this fell from ≈ 10 lux at the start of metamorphosis to between 1.0 and 10 lux at the end. The lower threshold light intensities in turbot compared to plaice may indicate that vision is of greater importance in the feeding of the former species.