Major Fouling Organisms Research Articles

Mode of action of antifouling compound albofungin in inhibiting barnacle larval settlement

Marine biofouling causes huge economic losses to the marine industry every year. Albofungin is a potential antifoulant showing strong anti-macrofouling activities against larval settlement of major fouling organisms. In the present study, directed RNA-seq and proteomic analyses were used to investigate changes in the transcriptome and proteome of a major fouling barnacle Amphibalanus amphitrite cyprids in response to albofungin treatment. Results showed that albofungin treatment remarkably upregulated the metabolism of xenobiotics by the cytochrome P450 pathway to discharge the compound and downregulated energy metabolic processes. Intriguingly, immunostaining and whole-mount insitu hybridization (WISH) revealed the spatial expression patterns of selected differentially expressed genes (glutathione S-transferase [GST], nitric oxide synthase [NOS], and calmodulin [CaM]) distributed in the thorax and antennule of A.amphitrite. Our study provides new insights into the mechanism of albofungin in interrupting the larval settlement of A.amphitrite and suggests its potential application as an antifouling agent in marine environments.

Biofouling characteristics in Xinghua Bay of Fujian, China

Biofouling is one of the main factors affecting the efficiency and safety of cooling water systems in coastal nuclear power plants. Understanding the population dynamics, succession rules and cumulative effects of major fouling organisms is the basis for targeted prevention and control. A 1-year simulated concrete panel test was conducted from December 2020 to November 2021 in Xinghua Bay, China. A total of 78 species of fouling organisms were recorded by combining the monthly, seasonal, semiannual, annual and monthly cumulative panels, and the community composition was dominated by nearshore warm-water species, making for a typical subtropical inner bay-type community. The fouling organisms had a peak attachment period from June to October. Significantly more attachment was observed during summer (from June to August) than during the other three seasons. The attachment amount in the second half-year (from June to November) was much higher than that in the first half-year (from December to May). The attachment thickness, density, and biomass of the bottom summer panels reached 20 cm, 105,150 ind./m2, and 19,274.50 g/m2, respectively, while those of the bottom annual panels were 40 cm, 27,300 ind./m2, and 17,762.50 g/m2, respectively. The dominant fouling organisms with calcified shells mainly included Amphibalanus reticulatus and Pernaviridis. These species had high attachment amounts,could accumulate attachments for a long time, and even might cause secondary blockage, making them the most detrimental to the safety of a cooling system. Moreover,the seasonal upward growth of hydroids and bryozoans can also significantly reduce the efficiency of cooling water intake. We suggest that targeted prevention and control should be carried out according to the larval attachment period of different dominant groups of fouling organisms during June-October, which can greatly improve the prevention and control efficiency. Strengthening the research on the biological cycle phenomenon of the main species and their main environmental impact factors, and establishing a scientific and effective early-warning model are the governance direction of formulating and implementing scientific pollution prevention and control in the future.

Discovery, Yield Improvement, and Application in Marine Coatings of Potent Antifouling Compounds Albofungins Targeting Multiple Fouling Organisms.

Marine biofouling caused huge economic losses of maritime industries. We aim to develop high-efficient, less-toxic, and cost-effective antifoulants to solve the problems of biofouling. In this study, we described the antifouling compounds albofungin and its derivatives (albofungin A, chrestoxanthone A, and chloroalbofungin) isolated from the metabolites of bacterium Streptomyces chrestomyceticus BCC 24770, the construction of high-yield strains for albofungin production, and application of albofungin-based antifouling coatings. Results showed that these albofungins have potent antibiofilm activities against Gram-positive and Gram-negative bacteria and anti-macrofouling activities against larval settlement of major fouling organisms with low cytotoxicity. With the best antifouling activity and highest yield in bacterial culture, albofungin was subsequently incorporated with hydrolyzable and degradable copolymer to form antifouling coatings, which altered biofilm structures and prevented the settlement of macrofouling organisms in marine environments. Our results suggested that albofungins were promising antifouling compounds with potential application in marine environments.

Constant exposure to environmental concentrations of the antifouling biocide Sea-Nine retards growth and reduces acetylcholinesterase activity in a marine mysid

Sea-Nine (4,5-dichloro-2-n-octyl-4-isothiazoline3-one; DCOIT) antifoulant has been widely used owing to its broad spectrum of biocide activity against major fouling organisms. In this study, several physiological parameters of a marine mysid were analyzed upon exposure to sublethal environmental concentrations (1 and 100 ng L−1) of Sea-Nine in two exposure conditions, intermittent (weekly; once per week) and constant (daily; once per 24 h) exposure, for 4 weeks. In both experimental conditions, growth retardation, acetylcholinesterase (AChE) activity, glutathione S-transferase (GST) activity, and number of newborn juveniles as second generation, together with their survival were measured. Morphometric parameters of total body, antennal scale, exopod, endopod, and telson were significantly retarded by 22%, 14%, 13%, and 24%, respectively, by daily exposure to 100 ng L−1 Sea-Nine for 4 weeks. Significant inhibition of AChE activity was observed at week 4 in the 100 ng L−1 daily Sea-Nine-exposed groups, whereas no significant GST activity was measured at the same experimental conditions. Inhibition of AChE activity would be associated with impairment of cholinergic system and may adversely modulate growth parameters of the mysid. The number of newly hatched juveniles from females that were exposed daily to 100 ng L−1 Sea-Nine was significantly lower than that of the control. Although no significant differences were observed between survival percentages of newborn juveniles for 30 days, mortality (NOEC and LC50) increased in the surviving offspring from the 100 ng L−1-exposed 1st generation of mysids. These findings suggested that constant exposure to Sea-Nine has detrimental effects on the growth parameters of marine mysids with inhibition of AChE activity.

Toward understanding barnacle cementing by characterization of one cement protein-100kDa in Amphibalanus amphitrite

Barnacles, as major fouling organisms, have attracted more attentions. It is no doubt that the study on cement proteins is required to illustrate the mechanism of barnacle cementing. A cement protein defined as Aa-cp100k was characterized from Amphibalanus amphitrite in this study. The amino acid sequence of Aa-cp100k was shown a high similarity to other three barnacles including Megabalanus rosa (Mr-cp100k), Tetraclita japonica formosana (Tj-cp100k) and Pollicipes pollicipes (Pp-cp100k). Moreover, the localization of Aa-cp100k in the vacuoles of cyprid β secretory cells and the adult cement gland cells by immunofluorescence microscopy, indicating that Aa-cp100k existed in both cyprid and adult barnacle. Aa-cp100k from basal plate could be dissolved in urea buffer without high concentration of dithiothreitol (DTT), different from that in Megabalanus rosa, implying diverse possible roles of cp100k in cementing.

Interactions between natural biofilm, substratum surface wettability, and mussel plantigrade settlement

Mytilus galloprovincialis is a major fouling organism in the inter-tidal zone. However, the interactions between M. galloprovincialis plantigrade settlement, biofilm characteristics, and surface wettability remains unknown. Here, we examined M. galloprovincialis plantigrade settlement responses to marine biofilms (BFs) on surfaces of varying wettability. No significant difference in mussel settlement was observed on young BFs (7 d) on surfaces of differing wettability; while settlement decreased on older BFs (14, 21, and 28 d) formed on low compared to high wettability surfaces. Surface wettability affected BF characteristics. The standardized harmonic mean and water contact angles values were not correlated with diatom density and chlorophyll a concentration, but were correlated with bacterial density, dry weight, and thickness. Denaturing gradient gel electrophoresis revealed that bacterial community structure differed on BFs on surfaces of varying wettability. Thus, surface wettability affects biofilm characteristics, and the subsequent changes in BF characteristics may be responsible for the variation in biofilm-inducing activity of M. galloprovincialis plantigrade settlement.

Antifouling Activity towards Mussel by Small-Molecule Compounds from a Strain of Vibrio alginolyticus Bacterium Associated with Sea Anemone Haliplanella sp.

Mussels are major fouling organisms causing serious technical and economic problems. In this study, antifouling activity towards mussel was found in three compounds isolated from a marine bacterium associated with the sea anemone Haliplanella sp. This bacterial strain, called PE2, was identified as Vibrio alginolyticus using morphology, biochemical tests, and phylogenetic analysis based on sequences of 16S rRNA and four housekeeping genes (rpoD, gyrB, rctB, and toxR). Three small-molecule compounds (indole, 3-formylindole, and cyclo (Pro-Leu)) were purified from the ethyl acetate extract of V. alginolyticus PE2 using column chromatography techniques. They all significantly inhibited byssal thread production of the green mussel Perna viridis, with EC50 values of 24.45 μg/ml for indole, 50.07 μg/ml for 3-formylindole, and 49.24 μg/ml for cyclo (Pro-Leu). Previous research on the antifouling activity of metabolites from marine bacteria towards mussels is scarce. Indole, 3-formylindole and cyclo (Pro-Leu) also exhibited antifouling activity against settlement of the barnacle Balanus albicostatus (EC50 values of 8.84, 0.43, and 11.35 μg/ml, respectively) and the marine bacterium Pseudomonas sp. (EC50 values of 42.68, 69.68, and 39.05 μg/ml, respectively). These results suggested that the three compounds are potentially useful for environmentally friendly mussel control and/or the development of new antifouling additives that are effective against several biofoulers.

First record of the non-native bryozoan Amathia (= Zoobotryon) verticillata (delle Chiaje, 1822) (Ctenostomata) in the Galápagos Islands

The warm water marine bryozoan Amathia (= Zoobotryon) verticillata (delle Chiaje, 1822) is reported for the first time in the Galapagos Islands based upon collections in 2015. Elsewhere, this species is a major fouling organism that can have significant negative ecological and economic effects. Comprehensive studies will be necessary to determine the extent of the distribution of A. verticillata in the Galapagos and its effects on native species. The establishment of A. verticillata emphasizes the need for regular monitoring of those vectors that could bring additional non-native species to the Archipelago.

Methods for the induction of reproduction in a tropical species of filamentous ulva.

The green seaweed Ulva is a major fouling organism but also an edible aquaculture product in Asia. This study quantified for the first time the effect of key factors on the reproduction of a tropical species of filamentous Ulva (Ulva sp. 3). The controlled timing of release of swarmers (motile reproductive bodies) was achieved when experiments were initiated in the early afternoon by exposing the thalli to a temperature shock (4°C) for 10 min and subsequently placing them into autoclaved filtered seawater under a 12 h light: 12 h dark photoperiod at 25°C. The release of swarmers then peaked two days after initiation. In contrast, segmentation, dehydration, salinity or time of initiation of experiments had no effect of any magnitude on reproduction. The released swarmers were predominantly biflagellate (95%), negatively phototactic and germinated without complementary gametes. This indicates that Ulva sp. 3 has a simple asexual life history dominated by biflagellate zoids.

Experimental studies on the effect of different metallic substrates on marine biofouling

In the wake of adoption of the resolution by the International Maritime Organization to control biofouling on vessels, which is recognized as a major vector for transfer of invasive species, this study attempts to create a baseline data on major hard-shelled biofouling organisms in the harbour waters. This study was primarily focused towards understanding the biofouling and corrosion pattern on various metals and their performance under immersed condition in a marine environment, at 0.3 and 3.0m depths. Furthermore, the study attempts to understand the surface dependent characteristics of barnacle base plate and its adhesion strength. Barnacle, mussels and oysters were the major fouling organisms accounting for 72.33% of the variation. Stainless steel and Titanium panels showed the highest average biofouling load of 176.36 and 168.35g/300cm2, respectively. The variance in biofouling between metals and depths was highly significant at p<0.001 and p<0.01, respectively. Morphology of barnacle base plate interfacial surface varied between metals. Barnacles with 8–9mm base diameter showed the maximum adhesion strength in shear of 6.86±0.95kPa.

Optimising settlement assays of pediveligers and plantigrades of Mytilus galloprovincialis

The mussel Mytilus galloprovincialis is a common aquaculture species, and also a major fouling organism that has negative economic impacts. There are no standard assay conditions for this important species and therefore, this study quantified the effect of key factors on the settlement of pediveligers and plantigrades. Density dependent settlement did not occur for either pediveligers or plantigrades. Settlement increased in drop assays in a 12 h light:12 h dark cycle, while bottom shade had no effect of any magnitude. In addition, settlement was significantly enhanced by storing pediveligers for between 4 and 24 days at 4°C. Overall, these data provide the template to optimise and standardise static laboratory settlement assays for mussels in order to develop materials that either enhance settlement for the aquaculture industry, or deter settlement for antifouling applications. Furthermore, simple mechanisms such as storage at 4°C can enhance settlement beyond current methods used in aquaculture hatcheries.

The effect of butenolide on behavioral and morphological changes in two marine fouling species, the barnacle Balanus amphitrite and the bryozoan Bugula neritina

Butenolide [5-octylfuran-2(5H)-one] is a very promising antifouling compound. Here, the effects of butenolide on larval behavior and histology are compared in two major fouling organisms, viz. cypris larvae of Balanus amphitrite and swimming larvae of Bugula neritina. Butenolide diminished the positive phototactic behavior of B. amphitrite (EC50 = 0.82 μg ml−1) and B. neritina (EC50 = 3 μg ml−1). Its effect on the attachment of cyprids of B. amphitrite was influenced by temperature, and butenolide increased attachment of larvae of B. neritina to the bottom of the experimental wells. At concentrations of 4 μg ml−1 and 10 μg ml−1, butenolide decreased attachment of B. amphitrite and B. neritina, respectively, but the effects were reversible within a certain treatment time. Morphologically, butenolide inhibited the swelling of secretory granules and altered the rough endoplasmic reticulum (RER) in the cement gland of B. amphitrite cyprids. In B. neritina swimming larvae, butenolide reduced the number of secretory granules in the pyriform-glandular complex.

Potent antifouling compounds produced by marine Streptomyces

Biofouling causes huge economic loss and a recent global ban on organotin compounds as antifouling agents has increased the need for safe and effective antifouling compounds. Five structurally similar compounds were isolated from the crude extract of a marine Streptomyces strain obtained from deep-sea sediments. Antifouling activities of these five compounds and four other structurally-related compounds isolated from a North Sea Streptomyces strain against major fouling organisms were compared to probe structure–activity relationships of compounds. The functional moiety responsible for antifouling activity lies in the 2-furanone ring and that the lipophilicity of compounds substantially affects their antifouling activities. Based on these findings, a compound with a straight alkyl side-chain was synthesized and proved itself as a very effective non-toxic, anti-larval settlement agent against three major fouling organisms. The strong antifouling activity, relatively low toxicity, and simple structures of these compounds make them promising candidates for new antifouling additives.

Physiological responses to hypoxia and anoxia in Balanus amphitrite (Cirripedia: Thoracica)

The barnacle Balanus amphitrite Darwin is a dominant intertidal and major fouling organism. As its life cycle includes both sessile and pelagic phases, adaptation is a necessity to toler- ate harsh conditions that are faced in the intertidal and pelagic environment. A downward trend in dissolved oxygen levels in coastal areas has been recently observed, which would have a more prominent effect on sessile organisms inhabiting the intertidal habitats. The effects of reduced dis- solved oxygen levels (hypoxia) and no oxygen (anoxia) on B. amphitrite adults and nauplii was assessed. Their response was evaluated by estimating the levels of the antioxidant enzymes catalase and superoxide dismutase. The impact of desiccation on antioxidant enzymes in adult B. amphitrite collected from different tidal heights was also evaluated. When nauplii were starved, 100% mortal- ity was observed after 64 h under hypoxia, and between 24 and 32 h under anoxia. However, when nauplii were exposed to hypoxia and anoxia along with food, their tolerance level increased. A decrease in naupliar feeding rate and oxygen consumption was observed when they were exposed to hypoxia and anoxia. A significant difference in the levels of antioxidant enzymes in adult B. amphitrite collected at different tidal levels was observed. Enzyme activity increased with increase in the duration of tidal exposure, suggesting a capability to adapt to stress in different niches. Nauplii showed antioxidant defense (increased enzyme activity) under hypoxia and anoxia. The magnitude of change in enzyme levels suggests that these enzymes could be potential biomarkers of stress.

Toxicity of crude extracts from several terrestrial plants to barnacle larvae on mangrove seedlings

This study involved the use of terrestrial plant extracts as substitutes for toxic chemicals to control the major fouling organisms of mangroves, the barnacles. In terms of the solvents used to obtain the plant crude extracts, ethyl acetate was best, ethyl alcohol next and pure water worst, except in the case of Allium cepa (AC), where pure water was better than ethyl alcohol. For different plants, different solvents resulted in different poisonous effects, and plants belonging to different classes showed different levels of activities. In particular, using ethyl acetate crude extracts, Allium sativum (AS), AC and Capsicum annuum (CA) caused massive death of barnacle larvae at a low concentration in the shortest time. The less poisonous organic solvents were able to extract more active materials, and this was advantageous in enhancing the effect. For pure water crude extracts, AC and Zingiber officinale were best, but for 95% ethyl alcohol crude extracts, AS and AC were best. For ethyl acetate crude extracts, the effects of AS, AC and CA were better. Different solvent extracts of AC with 12 h of exposure all had LC 50 values <100 mg/L. The concentrations of the effective plant crude extracts used were close to, or even lower than, those used for pesticides referred to in the literature.

Effects of initial surface wettability on biofilm formation and subsequent settlement of Hydroides elegans

Hydroides elegans is a major fouling organism in tropical waters around the world, including Pearl Harbor, Hawaii. To determine the importance of initial surface characteristics on biofilm community composition and subsequent colonization by larvae of H. elegans, the settlement and recruitment of larvae to biofilmed surfaces with six different initial surface wettabilities were tested in Pearl Harbor. Biofilm community composition, as determined by a combined approach of denaturing gradient gel electrophoresis and fluorescence in situ hybridization, was similar across all surfaces, regardless of initial wettability, and all surfaces had distinct temporal shifts in community structure over a 10 day period. Larvae settled and recruited in higher numbers to surfaces with medium to low wettability in both May and August, and also to slides with high wettability in August. Pearl Harbor biofilm communities developed similarly on a range of surface wettabilities, and after 10 days in Pearl Harbor all surfaces were equally attractive to larvae of Hydroides elegans, regardless of initial surface properties.

ムラサキイガイ幼生の着生機構

The mussel (Mytilus galloprovincialis) is an important seafood species and is cultivated in many countries. It is also considered a major fouling organism because it colonizes artificial structures causing economic problems. Many studies have demonstrated that M. galloprovincialis larval settlement is influenced by various substrata, i.e., macroalgae, conspecifics and biofilms. This review summarizes the roles of substrata on the larval settlement, with reference to the involvement of chemical cue(s). Literature on M. edulis larval settlement is also included. Larvae of M. galloprovincialis settle on specific filamentous algae in response to chemical cue(s), and not to the algal morphology or associated organisms. Evidence of the gregariousness in this species has also been demonstrated in the laboratory. Biofilms strongly induce the larval settlement and their activities are modulated by immersion periods and months (seasons). Bacteria in biofilms are directly involved in the induction of larval settlement and the viability of bacteria is a requirement for the production of chemical cue(s). Evidence also suggests that two chemical cues derived from bacteria, i.e., waterborne and surface-bound cues, act synergistically on the larval settlement. Further research on the identification of chemical cues is necessary to clarify the mechanism of mussel larval settlement.

沖縄の発電所におけるフジツボの付着と成長

We observed the settlement of barnacles on test panels from July, 1996, to March, 1999, at an electrical power generating station on the shore of Kin Bay in Okinawa Prefecture, Japan. Amphibalanus reticulatus and Tesseropora pacifica were the dominant species of barnacle on the test panels. Amphibalanus reticulatus settled mainly in April and from August to November. Tesseropora pacifica settled from September to January. Growth of A. reticulatus was slow in winter but became fast in spring. We also investigated the sessile organisms in the power station's water intake conduits, where barnacles proved to be among the major fouling organisms.

Thermal tolerance of the invasive oyster Crassostrea gigas: Feasibility of heat treatment as an antifouling option

Pacific oysters, Crassostrea gigas are traditionally considered shellfish of great fishery and aquaculture value. For these reasons they are introduced worldwide. Recently there has been increasing reports about the prevalence of C. gigas as biofouling organism in cooling water systems. In the absence of relevant data on the susceptibility of oysters to commonly employed antifouling techniques such as heat treatment, it was presumed that oysters would be controlled by treatment programmes directed against other major fouling organisms. The present study was carried out to test the above hypothesis, and results showed that C. gigas has an upper temperature tolerance that is much higher than other major marine fouling animals including blue mussel Mytilus edulis. Apparently, temperature regimes presently used in heat treatment of cooling water systems fouled by mussels need to be increased, if C. gigas are to be controlled effectively. Our results also indicate that previous exposure of C. gigas to sublethal high temperatures could make them more resistant to subsequent thermal treatment, an aspect that should be taken into account when heat treatment is used as a fouling control option against oyster fouling.

Seasonal dynamics of the numbers of larvae of common fouling organisms in Balaklava Bay (Black Sea)

The seasonal dynamics of the numbers of larvae of common fouling organisms—the barnacle Balanus improvisus and the bivalves Mytilaster lineatus and Mytilus galloprovincialis in Balaklava Bay (Black Sea) —were investigated during 2000 and 2001 within the framework of an ecological research project. Under the conditions of an increased anthropogenic load, seasonal fluctuations of the numbers and the distribution of larvae depend on rhythms of the breeding cycles of fouling invertebrates and on the hydrodynamic features of the region (water setup). The differences in the optimum temperatures, hatching intensity, and time of occurrence of larvae in the plankton, as well as the irregular distribution of larvae, allow the three major fouling organisms to reduce interspecific competition for food and the substrate.