Calcium is necessary for light excitation in barnacle photoreceptors

Summary Illumination of barnacle (Balanus amphitrite) photoreceptors is known to increase the membrane permeability to sodium and Ca²⁺ ions resulting in a depolarizing receptor potential. In this report, we show that lanthanum (La³⁺), a known inhibitor of Ca-binding proteins, reversibly eliminates the receptor potential of barnacle photoreceptors when applied to the extracellular space. Similar reversible elimination of the light response was obtained by removing extracellular Ca²⁺ by application of the calcium chelating agent EGTA. Iontophoretic injection of Ca²⁺, but not K⁺ into the cells protected both the transient and the steady-state phases of the receptor potential from elimination by EGTA while only the transient phase was protected in the presence of La³⁺. The EGTA experiments suggest that internal Ca²⁺ is necessary for light excitation of barnacle photoreceptors while the La³⁺ experiments suggest that La³⁺-sensitive inward current is necessary to maintain excitation during prolonged light.Abbreviations EGTA ethylenglyol-bis-(-aminoethylether) N, N, N¹, N¹-tetraacetate - BAPTA bis-(0-aminophenoxy)-ethane-N, N, N¹, N¹-tetraacetic acid - DMSO dimethyl sulfoxide - trp transient receptor potential - nss no steady state - ASW artificial sea water     

Stylochus (Imogene) matatasi n. sp. (Platyhelminthes,Polycladida): pest of cultured giant clams and pearl oysters from Solomon Islands

A large polyclad flatworm has been consistently found associated with mortalities of the cultured giant clam,Tridacna gigas (L.) and the fouling pearl oysterPinctada maculata (Gould) in Solomon Islands.Stylochus (Imogene) matatasi n. sp. is described and a brief account of its biology is given.ICLARM Contribution # 856.ICLARM Contribution # 856.     

Immunocytochemical studies on the naupliar nervous system of Balanus improvisus (Crustacea, Cirripedia, Thecostraca)

The nervous system of nauplii of the crustacean taxon Cirripedia was analysed in the species Balanus improvisus Darwin, 1854 using for the first time immunocytochemical staining against serotonin, RFamide and α-tubulin in combination with confocal laser scanning microscopy. This approach revealed a circumoesophageal neuropil ring with nerves extending to the first and second antennae and to the mandibles, all features typical for Crustacea. In addition, RFamidergic structures are present in the region of the thoraco-abdomen. A pair of posterior nerves and a pair of lateral nerves run in anterior-posterior direction and are connected by a thoracic nerve ring and a more posteriorly situated commissure. A median nerve is situated along the ventral side of the thoraco-abdomen. The innervation of frontolateral horns and the frontal filaments are α-tubulin-positive. Several pairs of large neurons in the protocerebrum, along the circumoesophageal connectives and in the mandibular ganglion stain only for serotonin. Due to the almost complete absence of comparable data on the neuroanatomy of early (naupliar) stages in other Crustacea, we include immunocytochemical data on the larvae of the branchiopod, Artemia franciscana Kellogg, 1906 in our analysis. We describe several characteristic neurons in the brains of the nauplius larvae of both species which are also found in decapod larvae and in adult brains of other crustaceans. Furthermore, our data reveal that the naupliar brain of cirripedes is more complex than the adult brain. It is concluded that this ontogenetic brain reduction is related to the sessile life style of adult Cirripedia.     

Evidence for different pharmacological targets for imidazoline compounds inhibiting settlement of the barnacle Balanus improvisus

We describe the effect of eight different imidazoline/guanidinium compounds on the settlement and metamorphosis of larvae of the barnacle Balanus improvisus. These agents were chosen on the basis of their similar pharmacological classification in vertebrates and their chemical similarity to medetomidine and clonidine, previously described as highly potent settlement inhibitors (nanomolar range). Seven of the tested compounds were found to inhibit settlement in a dose-dependent manner in concentrations ranging from 100 nM to 10 microM without any significant lethal effects. In vertebrate systems these substances have overlapping functions and interact with both alpha-adrenoceptors as well as imidazoline binding sites. Antagonizing experiments using the highly specific alpha(2)-antagonist methoxy-idazoxan or agmatine (the putative endogenous ligand at imidazoline receptors) were performed to discriminate between putative pharmacological mechanisms involved in the inhibition of cyprid settlement. Agmatine was not able to reverse the effect of any of the tested compounds. However, methoxy-idazoxan almost completely abolished the settlement inhibition mediated by guanabenz (alpha(2)-agonist, I(2) ligand), moxonidine (alpha(2)-agonist, I(1) ligand) and tetrahydrozoline (alpha-agonist, I(2) ligand). The actions of cirazoline (alpha(1)-agonist, I(2) ligand) BU 224 (I(2) ligand) and metrazoline (I(2) ligand) were not reversed by treatment with methoxy-idazoxan. These results suggest that the settlement inhibition evoked by the I(2) ligands and alpha(2)-agonists used in this study of the neurologically simple but well-organized barnacle larva is mediated through different physiological targets important in the overall settlement process.     

Influence of predator density on nonindependent effects of multiple predator species

Interactions between multiple predator species are frequent in natural communities and can have important implications for shared prey survival. Predator density may be an important component of these interactions between predator species, as the frequency of interactions between species is largely determined by species density. Here we experimentally examine the importance of predator density for interactions between predator species and subsequent impacts on prey. We show that aggressive interactions between the predatory shore crabs Carcinus maenas and Hemigrapsus sanguineus increased with predator density, yet did not increase as fast as negative interactions between conspecifics. At low density, interactions between conspecific and heterospecific predators had similar inhibitory impacts on predator function, whereas conspecific interference was greater than interference from heterospecifics at high predator density. Thus the impact of conspecific interference at high predator density was sufficient in itself that interactions with a second predator species had no additional impact on per capita predation. Spatial and temporal variability in predator density is a ubiquitous characteristic of natural systems that should be considered in studies of multiple predator species.     

Geocoris punctipes as a predator of Bemisia tabaci: a laboratory evaluation

Geocoris punctipes (Say), a predaceous lygaeid not previously docmented as a whitefly predator, was tested in the laboratory as a natural enemy of the sweet potato whitefly, Bemisia tabaci (Genn.). Its stalking behavior with whiteflies as prey was similar to that observed with aphids. A previously unobserved behavior was noted that involves the predators using salivary secretions to fasten the wings of prey to various surfaces, allowing labial probing and feeding. Prey consumption as a function of prey number appeared to follow the pattern of the Holling type II functional response. Handling time per prey item ranged from about 180 to 240 seconds. No changes were observed in handling time devoted to earlier versus later catches. Nutritional quality of whiteflies was measured using crude protein, lipids and carbohydrates as criteria. Performance, in terms of predator behavior, total daily handling time, functional response, energy budget and nutritional quality all support the hypothesis that G. punctipes is a promising candidate for biological control of sweet potato whiteflies.     

Biological studies of the flesh flySarcophaga (Parasarcophaga) misera and its effects as a predator of the snailIndoplanorbis exustus

Attempts were made to identify a natural predator of the freshwater snailIndoplanorbis exustus, vector of schistosomiasis. Ochterlony's double immunodiffusion technique was used. The flySarcophaga (Parasarcophaga) misera was identified as a predator ofI. exustus. Various aspects of the biology of this fly and its effects as a predator of snailI. exustus were studied in the laboratory. The possible use of the larvae of this fly to controlI. exustus is discussed as an additional control measure to chemical control.      

Distribution and abundance of a nemertean egg predator ( Carcinonemertes sp.) on a leucosiid crab,Randallia ornata

The purple globe crab, Randallia ornata (Randall, 1839) (Decapoda: Leucosiidae) is a common crustacean found on sandy beaches from Northern California to Baja California, Mexico. An undescribed species of Carcinonemertes, which shares biological features with a partially described worm from Tasmania was recovered from R. ornata. The worm reaches 10 mm in length and secretes a sturdy tube with a distinctive spiral shape. The total prevalence of the worm on R. ornatawas 70% (85% for ovigerous female crabs, 67% for post-ovigerous female crabs, and 45% for male crabs). Intensities reached 32 worms per crab. Adult worms were found in crab egg masses, regressed adults were recovered from under the abdomen of post-ovigerous females, and encysted juveniles were found on the gill lamellae of non-ovigerous female crabs and occasionally from the gill lamellae of male crabs.     

Rearing of a coleopterous predator,Trogossita japonica (Col.: Trogossitidae), on artificial diets

Artificial diets for a coleopterous predator of pine bark and wood borers,Trogossita japonica Reitter (Col.: Trogossitidae) were studied. Diets composed of silkworm pupa-powder, dry yeasts, sucrose, peptone, squid liver oil, agar and distilled water supported larval development. The size of diet-reared adults was smaller than that in the control but they mated and oviposited normally when maintained on prey insects. An aseptic rearing method for the larvae of predator, that required a single diet change, was devised.

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Résumé Nous avons élaboré et évalué quatre milieux semi-synthétiques pour l'élevage deTrogossita japonica Reitter, prédateur d'insectes sous-corticaux et xylophages des pins. Les larves ont été nourries avec des milieux composés de poudre de chrysalide du ver à soie, levure de bière séchée, sucrose, peptone, huile de foie de seiche, agar et eau distillée. Les adultes obtenus ont été plus petits que ceux du lot témoin, mais ils se sont accouplés et ont pondu normalement quand ils ont été nourris avec des proies-insectes. Une méthode d'élevage des larves sur milieu aseptique a été élaborée, laquelle ne nécessite qu'un changement de milieu nutritionnel au cours du développement larvaire du prédateur.

     

Some effects of early starvation on the survival and development of barnacle nauplii,Balanus improvisus (Darwin)

Newly hatched stage I–II nauplii of Balanus improvisus (Darwin) were “totally starved” (until death) or “partially starved” for the first 48 h and 96 h of their development. Daily mortality and molting were monitored throughout larval development in both starved and fed control groups. Fed control animals exhibited a largely synchronous molting pattern with instars of equal duration. Total starvation suppressed molting beyond stage II; 50% mortality occurred in ≈4 days at both 15 and 21 °C, while longest survival time was 7 days at 15 °C and 6 days at 21 °C. At 15 °C, partially starved nauplii retained the ability to complete naupliar development but at a slower overall rate and with increased mortality relative to controls. These effects were more pronounced in the 96-h group. Increased mortality of stage VI nauplii was evident in both partially starved groups (7.1% for 48 h, 18.8% for 96 h) relative to unstarved controls (3.1%). Stage II nauplii exhibited little resistance to starvation and survival potential may have decreased as soon as 24 h.     

Testing the threat-sensitive predator avoidance hypothesis: physiological responses and predator pressure in wild rabbits

Predation is a strong selective force with both direct and indirect effects on an animal’s fitness. In order to increase the chances of survival, animals have developed different antipredator strategies. However, these strategies have associated costs, so animals should assess their actual risk of predation and shape their antipredator effort accordingly. Under a stressful situation, such as the presence of predators, animals display a physiological stress response that might be proportional to the risk perceived. We tested this hypothesis in wild European rabbits (Oryctolagus cuniculus), subjected to different predator pressures, in Doñana National Park (Spain). We measured the concentrations of fecal corticosterone metabolites (FCM) in 20 rabbit populations. By means of track censuses we obtained indexes of mammalian predator presence for each rabbit population. Other factors that could modify the physiological stress response, such as breeding status, food availability and rabbit density, were also considered. Model selection based on information theory showed that predator pressure was the main factor triggering the glucocorticoid release and that the physiological stress response was positively correlated with the indexes of the presence of mammalian carnivore predators. Other factors, such as food availability and density of rabbits, were considerably less important. We conclude that rabbits are able to assess their actual risk of predation and show a threat-sensitive physiological response.     

Molecular Detection of Marine Invertebrate Larvae

The ecological patterns of many invertebrate larvae remain an ongoing mystery, in large part owing to the difficult task of detecting them in the water column. The development of nucleic-acid–based technology has the potential to resolve this issue by direct identification and monitoring of embryonic and larval forms in situ. We report herein on the successful development and application of nucleic-acid–based sandwich hybridization assays that detect barnacles using rRNA-targeted probes with both group-(order Thoracica) and species-(Balanus glandula) specificity. Primary results include the determination of target 18S rRNA sequences and the construction of “capture” probes for detection of larvae using hybridization techniques. In addition, we modified existing protocols for whole cell hybridization of invertebrate larvae as confirmation of the sandwich hybridization results. We used both hybridization techniques successfully in the laboratory on a plankton time series collected over 3 months, as well as a week-long in situ deployment of the technique in Monterey Bay, CA. The adaptability of this technology promises to be further applicable to various organisms and could be used to enhance our understanding of larval presence in the world's oceans.     

Variability of Acartia clausi in the Black Sea

There are two morphotypes of A. clausi in the Black Sea. The diversity in the spine ornamentation of the last prosomal and urosomal segments generally has been decreasing in both morphotypes during the period of this study (1976–1990). Seasonal data showed increased variability in April.     

Acartia tonsa: a species new for the Black Sea fauna

The presence of Acartia tonsa in Black Sea waters (South Crimean coast, Ukraine) is reported for the first time. A. tonsa may be a recently introduced species in the Black Sea. Morphological features of the adults have been shown by S.E.M.     

Emergent multiple predator effects in an experimental microbial community

Studies on multiple predator effects have typically focused on metazoans. Here we examined the effects of multiple consumers on bacteria. Specifically, we investigated the separate and combined effects of two common bacterivorous ciliates, Colpidium striatum and Paramecium caudatum, on a simple bacterial assemblage. Non-additive multiple predator effects emerged for all bacterial species significantly affected by grazing, where bacterial responses in the two-consumer treatment cannot be predicted by summing the separate effects of the two consumers. Some species showed risk reduction (observed responses less than expected), whereas others showed risk enhancement (observed responses larger than expected). We attributed risk alteration to the interference between the two consumers. Contradictory to theoretical predictions, total bacterial abundance in the two-consumer treatment did not differ from single-consumer treatments and consumer-free controls, due largely to risk reduction and compensatory responses within the bacterial assemblage. Decomposition of particulate organic matter was greater in the two-consumer treatment than the Paramecium single-consumer treatment, but did not differ among other treatments. These results suggest that the presence of multiple consumers may have unexpected impacts on the structure and functioning of bacterial communities.     

Multi-seasonal barnacle (Balanus improvisus) protection achieved by trace amounts of a macrocyclic lactone (ivermectin) included in rosin-based coatings

Rosin-based coatings loaded with 0.1% (w/v) ivermectin were found to be effective in preventing colonization by barnacles (Balanus improvisus) both on test panels as well as on yachts for at least two fouling seasons. The leaching rate of ivermectin was determined by mass-spectroscopy (LC/MS-MS) to be 0.7 ng cm^?2 day^?1. This low leaching rate, as deduced from the Higuchi model, is a result of the low loading, low water solubility, high affinity to the matrix and high molar volume of the model biocide. Comparison of ivermectin and control areas of panels immersed in the field showed undisturbed colonisation of barnacles after immersion for 35 days. After 73 days the mean barnacle base plate area on the controls was 13 mm², while on the ivermectin coating it was 3 mm². After 388 days, no barnacles were observed on the ivermectin coating while the barnacles on the control coating had reached a mean of 60 mm². In another series of coated panels, ivermectin was dissolved in a cosolvent mixture of propylene glycol and glycerol formal prior to the addition to the paint base. This method further improved the anti-barnacle performance of the coatings. An increased release rate (3 ng cm^?2 day^?1) and dispersion of ivermectin, determined by fluorescence microscopy, and decreased hardness of the coatings were the consequences of the cosolvent mixture in the paint. The antifouling mechanism of macrocyclic lactones, such as avermectins, needs to be clarified in further studies. Beside chronic intoxication as ivermectin is slowly released from the paint film even contact intoxication occurring inside the coatings, triggered by penetration of the coating by barnacles, is a possible explanation for the mode of action and this is under investigation.     

Prey preference and feeding capacity of the larvae ofAblattaria arenaria [Coleoptera: Silphidae], a snail predator

The larval stages ofAblattaria arenaria were provided with 4 different snail species:Monacha syriaca (Ehrenberg),Xeropicta derbentina (Krynicki),Candidula sp., andZebrina eburnea (Pfeiffer) to determine if the prey species affected developmental time and food preference of larvae. Functional response of each larval stage ofA. arenaria was also tested for increasing density ofX. derbentina, the most common prey species found in association withA. arenaria locally. The developmental time of each larval stage did not show any statistical difference when fed with different snail species. The total developmental time from egg hatch to adult emergence was 19.0, 19.1, 18.0 and 21.4 days for prey speciesM. syriaca, X. derbentina, Candidula sp., andZ. eburnea, respectively. When prey was offered to larvae either as a single species or as combination of several species,M. syriaca was the most preferred. The prey least consumed wasCandula sp. when prey was given separately, andZ. eburnea was least preferred when other prey species were present in the arena. The 3^rd larval stage did not eat anyZ. eburnea if other prey species were present. The amount of prey consumed by the 1^st larval stage did not show any statistical differences with increasing density ofX. derbentina. But the response of 2^nd and 3^rd larval stages was very similar to each other although the amount of prey they consumed was very different. They both showed a rapid increase in consumption rate at early densities, then a negatively but slowly accelerated rise to plateaus at higher densities, a type-2 functional response curve. All larval stages were very sensitive to starvation. Mortality started after the 2^nd day, and all individuals of all larval stages were dead by the 5^th day.      

Host plants affect predator fitness via the nutritional value of herbivore prey: Investigation of a plant-aphid-ladybeetle system

The interactions among host plants(Medicago sativa L., cv. `OKO8' and Vicia faba L., cv. `Windsor'), aphid prey(Acyrthosiphon pisum Harris, Homoptera:Aphididae), and Coccinella septempunctata L. (Coleoptera: Coccinellidae) preimaginal biology were evaluated. Interactions were measured over a range of limiting daily prey levels (1.2 mg–16.4 mg)from each host plant colony. Compared withA. pisum reared on V. faba, A. pisum reared on M. sativa storedsignificantly more fatty acids which resultedin a 1.17-fold increase in available caloriesfor developing C. septempunctata. Theincreased survival, decreased developmentaltimes, and larger size of C.septempunctata supplied with A. pisumreared on M. sativa clearly demonstratehost plant effects at the third trophic level. At low very limiting daily prey levels, A. pisum reared on M. sativa were moresuitable prey for C. septempunctatasurvival, development, and adult size thanA. pisum reared on V. faba. Coccinella septempunctata survival ratios(larval), developmental times, and adult sizeconverged (were not statistically different)between host plants at higher daily A.pisum levels. These convergence's supportthe hypothesis that there were quantitativedifferences in the nutritional value ofaphids, as influenced by differences in fattyacids and subsequent nutritional levels(calories), between aphids reared on separateplant hosts. The observed tritrophicinteractions appear to be modulated by thebiochemical response of A. pisum to hostplants.     

Oviposition choices by a pre-dispersal seed predator (Hylemya sp.)

Summary Pre-dispersal seed predators must choose proper hosts and flowers on those hosts that have a high probability of setting seed. In this study, I documented that flowers of Ipomopsis aggregata (Polemoniaceae) chosen by a pre-dispersal seed predator (Anthomyiidae: Hylemya sp.) had a higher probability of seed set than flowers the fly did not choose. I experimentally tested the hypothesis that Hylemya uses a post-fertilization cue in making oviposition choices. I found no evidence for this hypothesis.     

Notes on the biology of some psammophile Turbellaria of the Black Sea

The biology of two marine turbellarians, Cercyra hastata (Tricladida) and Pseudomonocelis ophiocephala (Proseriata) has been studied over a period of five years. They are the main components of the biocoenosis of the saccocirrus sand of Sevastopol Bay. These species have a significant role in the processes of secondary production and transformation of the organic matter in the coastal zone.