The impact and control of biofouling in marine aquaculture: a review.

Biofouling in marine aquaculture is a specific problem where both the target culture species and/or infrastructure are exposed to a diverse array of fouling organisms, with significant production impacts. In shellfish aquaculture the key impact is the direct fouling of stock causing physical damage, mechanical interference, biological competition and environmental modification, while infrastructure is also impacted. In contrast, the key impact in finfish aquaculture is the fouling of infrastructure which restricts water exchange, increases disease risk and causes deformation of cages and structures. Consequently, the economic costs associated with biofouling control are substantial. Conservative estimates are consistently between 5-10% of production costs (equivalent to US$ 1.5 to 3 billion yr(-1)), illustrating the need for effective mitigation methods and technologies. The control of biofouling in aquaculture is achieved through the avoidance of natural recruitment, physical removal and the use of antifoulants. However, the continued rise and expansion of the aquaculture industry and the increasingly stringent legislation for biocides in food production necessitates the development of innovative antifouling strategies. These must meet environmental, societal, and economic benchmarks while effectively preventing the settlement and growth of resilient multi-species consortia of biofouling organisms.

Making heads from tails: development of a reversed anterior-posterior axis during budding in an acoel.

The anterior-posterior axis is a key feature of the bilaterian body plan. Although axis specification during embryogenesis has been studied extensively, virtually nothing is known about how this axis can be established post-embryonically, as occurs in budding animals. We investigated bud formation in the acoel Convolutriloba retrogemma, which reproduces by a remarkable process involving the formation of animals with linked but completely opposite body axes. Reverse axes are established anew during each round of budding and manifestations of the bud's new axis develop gradually, with regionalization of axial patterning genes (Hox and otx) and the establishment of organized musculature occurring secondarily, after bud initiation. A swath of tissue at the parent-bud boundary has no regenerative potential and appears devoid of inherent axial polarity. GSK-3 inhibitor trials suggest that Wnt/beta-catenin or Hedgehog signalling may mediate the establishment of this unpolarized zone. Formation of unpolarized tissue may provide a buffer between opposing polarity cues and be a general mechanism by which budding animals establish and maintain linked body axes. In addition to elucidating the developmental basis of budding in a bilaterian, this study provides insight into convergence in animal budding mechanisms, redeployment of embryonic gene expression during budding, and Hox gene evolution.

Effects of neuropeptide F on regeneration in Girardia tigrina (Platyhelminthes).

The effects of neuropeptide F (NPF; from Moniezia expansa) on the regeneration of Girardia tigrina were studied. The animals were decapitated and incubated in water (control) or NPF. The dynamics of the proliferation of the neoblasts in the developing tissue were studied during the course of regeneration by monitoring the mitotic index (MI). The effects of incubation in FMRFamide and GYIRFamide on the MI were also tested. The course of cephalic regeneration was followed with in vivo computer-assisted morphometry for up to 7 days. The development of the regenerating nervous system and the musculature was visualised by immunostaining with a primary antiserum to the C-terminal decapeptide of NPF (YFAIIGRPRFa) and tetramethylrhodamine-isothiocyanate-conjugated phalloidin, which stains F-actin in muscle filaments. The study showed that NPF had a stimulatory effect on the mitotic activity of the neoblasts. FMRFamide and GYIRFamide did not have this effect. NPF also stimulated the growth of the regenerating head and the growing nervous system and musculature. NPF is postulated to have a morphogenetic action in the regenerating animals.

Responses of periphyton and invertebrates to a tetradecyl-pentadecyl sulfate mixture in stream mesocosms.

Alkyl sulfates (AS) are anionic surfactants widely used in household and personal cleansing applications. Aquatic toxicity of AS under laboratory conditions indicated effects at relatively low concentrations (50-230 microg/L) for some sensitive species. A comprehensive stream mesocosm study of an AS mixture composed of tetra- (C14) and pentadecyl (C15) chain lengths was conducted to better understand effects on microbial and macroinvertebrate populations and communities. A 56-d exposure of AS was performed at concentrations ranging from 57 to 419 microg/L (analytically confirmed exposures) and was accompanied by detailed investigations of periphyton community function (autotrophy, heterotrophy, and metabolism of test chemical), periphyton structure (algal population and community dynamics based on taxonomic identity), and invertebrate structure (benthic abundance, drift, and insect emergence patterns based on taxonomic identity). A no-observed-effect-concentration (NOEC) of 222 microg/L was concluded for several individual algal and invertebrate species based on univariate statistical analyses. An apparent energetic subsidy from C14-15AS at the highest concentrations of 222 to 419 microg/L was observed and tied to changes in microbial community processing of AS when added at these high concentrations. A multivariate analysis based on principal response curves (PRC) indicated that communities in streams exposed to 222 to 419 microg/L were significantly different from the controls leading to an overall (multivariate and univariate) conclusion that 106 microg/L was the ecosystem NOEC. Exposure to AS in the environment has been demonstrated to be in the range of 5 to 21 microg/L in 100% wastewater treatment plant effluent. Potential environmental effects are at least 5 to 20 times above worst-case environmental exposures; therefore, C14-15AS does not pose a risk to the aquatic environment due to normal use patterns.

A comparison of selected diversity, similarity, and biotic indices for detecting changes in benthic-invertebrate community structure and stream quality.

Implementation of advanced wastewater treatment at the two municipal wastewater-treatment plants for Indianapolis, Indiana, resulted in substantial improvement in the quality of the receiving stream and significant changes in the benthic-invertebrate community. Diversity, similarity, and biotic indices were compared to determine which indices best reflected changes in the composition of the biota in the river. None of the indices perfectly reflected the changes in river quality or community structure. Similarity indices, especially percentage similarity, exhibit the most promise of the three classes of indices. Diversity indices were least useful, wrongly indicating that water quality deteriorated after the upgrade of the wastewater-treatment plants. The most descriptive tool in analyzing the data was the percentage of Ephemeroptera, Plecoptera, and Trichoptera (EPT) taxa present. Using a mixture of indices and other analytical tools, such as EPT, in the analysis of biological data will ensure the most effective investigations of water quality.

Persistent effects of a serotonin depletor (p-chlorophenylalanine) in regenerated planaria (Dugesia dorotocephala).

p-Chlorophenylalanine (pCPA) caused an increase in looping and locomotion speed in planaria (Dugesia dorotocephala). These effects dissipated quickly after treatment, except in animals that regenerated from worms surgically sectioned after treatment. Planaria that regenerated from head sections showed more looping 6 and 11 days later than tail regenerates, regenerates from untreated worms, and uncut worms. After the initial acceleration, pCPA animals, especially regenerates from tail sections, slowed down compared with untreated controls. Tail regenerates showed reduced speed 6 and 11 days later in comparison with head regenerates, regenerates from untreated animals, and uncut animals. The speed effect may have been due to pathogenesis in the eye spots in pCPA animals. The looping effect, however, may have been due to persisting serotonin depletion in the head regenerates. It is suggested that research on this problem might profitably be pursued with sexually reproducing planaria, to examine the possibility of genetic recoding due to serotonin depletion.

A pharmacological study of dopaminergic receptors in planaria.

The dopaminergic receptors of planaria have been studied with pharmacological and biochemical criteria. Dopamine D1 selective agonists (CY 208243 (10 micrograms/ml) and SKF 38393 (10 micrograms/ml] induced in planaria typical screw-like hyperkinesias, that were inhibited by a D1 antagonist (SCH 23390 (10 micrograms/ml], but not by a D2 antagonist (sulpiride (1000 micrograms/ml]. Dopamine D2 selective agonists (PHNO (5 micrograms/ml), lisuride (5 micrograms/ml] on the contrary induced a typical "C" like curling, that was inhibited by pretreatment with D2 selective blocking agents, but not by D1 selective blocking agents. With agonists with a D1/D2 mixed action (apomorphine 60 micrograms/ml) or with amphetamine (100 micrograms/ml), the D1 type movements appeared to be more evident. Dopamine D1-selective agonists, mixed action agonists or D2-selective agonists, all induced a significant increase in levels of cAMP, that was prevented by pretreatment with the specific DA blocking agent.

Neoplastic transformation in the planarian: I. Cocarcinogenesis and histopathology.

Although several investigators have reported that exposure to mammalian carcinogens induces abnormal tumorlike growths and teratogenic remodeling in planarians, there is no general agreement that these, or comparable responses in any other invertebrates, model mammalian carcinogenesis. To investigate this question, freshwater planarians of the species Dugesia dorotocephala were exposed to culture water containing an initiator and a promoter, either alone or in combination. Cadmium, a potent carcinogen, was used as an initiator in the protocol. Treatment with sublethal concentrations of cadmium sulfate produced a benign, but persistent, tumor in a small percentage of the planarians. The addition of 12-O-tetradecanoylphorbol-13-acetate (TPA), a phorbol ester and well-known promoter, to the cadmium-containing solutions resulted in the induction of a progressive, potentially lethal, transplantable tumor in a large proportion of the treated flatworms. Light and electron microscopy revealed this particular tumor to be composed both of immature cells and of a single mature cell type: newly differentiated, but transformed, reticular cells. Further examination of the infiltrating tissue formations elucidated the profile of differentiation, from a population of mitotically active transformed stem cells through the transitional stages in the associated reticuloma. These results suggest that 1) the freshwater planarian displays the major phenomenology of mammalian cocarcinogenesis and that 2) the planarian reticuloma models several important features of a neoplastic stem cell disease.

Neoplastic transformation in the planarian: II. Ultrastructure of malignant reticuloma.

Cadmium and phorbol ester induced tumorigenesis in the planarian, Dugesia dorotocephala, develops as a cocarcinogenic process involving initiation and promotion in the progression of neoplastic disease. Treatment of intact planarians with sublethal concentrations of cadmium sulfate and 12-O-tetradecanoylphorbol-13-acetate (TPA) induced a type of infiltrating tumor that proved to be potentially lethal. Surgical transplantation of such tumorous tissues into otherwise healthy planarians resulted in the same histopathological progression to lethality, which confirmed the metastatic nature of the neoplasia. Electron microscopic studies revealed that both the chemically-induced and the transplantation-based tumors involved, exclusively, the proliferation and differentiation of abnormal reticular cells, referred to as reticuloma cells. Reticular cells normally are ameboid, phagocytic, and are thought to provide the planarian with a phylogenetic predecessor of an immune surveillance system. A considerable incidence of mitosis was observed within the tumor areas; and the sequence of differentiation, from transformed stem cells to mature but nonfunctional reticuloma cells, was elucidated. This profile of differentiation supports the concept of cellular derivation via stem cell dynamics as opposed to dedifferentiation. A variety of ultrastructural abnormalities were characterized: several of which tend to substantiate the anaplastic quality of the reticuloma, while others are more specifically diagnostic for malignancy. These findings further extend the potential usefulness of the planarian malignant reticuloma as a model system for the study of neoplastic stem cell diseases.

Stimulation of cellular proliferation and differentiation in the intact and regenerating planarian Dugesia(G) tigrina by the neuropeptide substance P.

The neuropeptide substance P (SP) is shown to be a potent mitogen for intact and regenerating planarians. At nanomolar concentrations, SP markedly enhances cellular proliferation causing an increase in the mitotic index and in the number of blastema cells. Moreover, albeit to a lower extent, SP enhances cellular differentiation as shown by the increases in eye and pharynx length in regenerating organisms. On the basis of these observations, we hypothesize that SP may be one of the postulated growth factors necessary for the stimulation of proliferation, and to a lesser extent differentiation of cells in intact and regenerating planarians.

Phylogenetic changes in sensitivity to Anemonia sulcata toxin (ATX II), and impact of first interaction with the toxin (imprinting) on later response to it.

The Anemonia sulcata toxin ATX II is cardiotoxic and neurotoxic, and--at a high dose level--even lethal for the mouse, neurotoxic, but non-lethal for the frog, and has no adverse influence whatever on the Planaria and Tetrahymena; it even stimulates the growth of the Tetrahymena at a low dose level. It also induces imprinting in the Tetrahymena, as judged from the altered response of the latter to ATX II on re-exposure. No similar imprinting effect was demonstrable in mice.

Planarians as a model system for in vitro teratogenesis studies.

Free-living flatworms such as planarians are inexpensive to culture, maintain, and use for toxicologic testing in the laboratory. A considerable number of basic studies by ourselves and others indicate that, in simplified miniature, they possess many features of biochemical and physiologic organization similar to higher animals such as mammals. These include a well-developed brain with a varied behavioral repertoire including complex maneuvers of prey capture and learning, with a number of the same neurotransmitters used in mammalian brain. They are sensitive to a variety of the same toxicants. Undifferentiated totipotent stem cells, i.e., "neoblasts," which are capable of mitosis and differentiation into any of the various specialized cell types, permit regeneration of complete planarians from fragments. They also provide new cells to replace those lost in the normal cellular turnover of nonregenerating planarians. Both regeneration of surgical fragments and aberrant remodeling of whole planarians model important features of embyrogenesis and are potentially useful for assaying teratogens. Results are described from studies in which various representative teratogenic toxicants were tested in these two different planarian paradigms. The potential of planarian cephalic regeneration for behavioral teratogenesis investigations is also indicated.

Inhibition of Dugesia tigrina auricle regeneration by inhibitors of polyamine synthesis.

Results are presented that indicate polyamine synthesis inhibitors increase the flatworm Dugesia tigrina's auricle regeneration time. This study serves as evidence that endogenous putrescine, spermidine, and spermine are necessary for the flatworm regeneration process.

The effect of sympathicomimetic agents on carbohydrate metabolism of planaria.

Epinephrine, ephedrine, dopamine and isoproterenol considerably influence carbohydrate utilization in planaria. Dopamine alone and in combination is the most potent, while ephedrine the least effective in this respect. The finding of increased responses to combined treatment supports the view of the existence of different receptor sites for the sympathicomimetic drugs. The results substantiate conclusions concerning the phylogenesis of hormone receptors.

Analysis of the progress of hormone-receptor amplification during differentiation in regenerating planarians.

Planarians developing from isolated regenerates, when pretreated with epinephrine at different periods of their second regeneration, show a higher rate of sugar uptake than the controls, which were not treated with epinephrine. The hormone receptor development can be influenced most effectively by pretreatments performed during the 1--3 and 6--9 day periods, though some effect can be evoked at any time in the course of the second regeneration. The head regenerates developing from the anterior end of the original midpiece (and regenerating tails the second time, B1) were found to be most sensitive, while the similar regenerates developing from the hindpiece of the original planarian (C) showed the weakest reactivity. The experiments provide further evidence that the hormone receptors' activity can be amplified in the course of regenerative development, leading thus--at a later stage--to a more intense sugar uptake response of the cells.