Increased inter-colony fusion rates are associated with reduced COI haplotype diversity in an invasive colonial ascidian Didemnum vexillum

Considerable progress in our understanding of the population genetic changes associated with biological invasions has been made over the past decade. Using selectively neutral loci, it has been established that reductions in genetic diversity, reflecting founder effects, have occurred during the establishment of some invasive populations. However, some colonial organisms may actually gain an ecological advantage from reduced genetic diversity because of the associated reduction in inter-colony conflict. Here we report population genetic analyses, along with colony fusion experiments, for a highly invasive colonial ascidian, Didemnum vexillum. Analyses based on mitochondrial cytochrome oxidase I (COI) partial coding sequences revealed two distinct D. vexillum clades. One COI clade appears to be restricted to the probable native region (i.e., north-west Pacific Ocean), while the other clade is present in widely dispersed temperate coastal waters around the world. This clade structure was supported by 18S ribosomal DNA (rDNA) sequence data, which revealed a one base-pair difference between the two clades. Recently established populations of D. vexillum in New Zealand displayed greatly reduced COI genetic diversity when compared with D. vexillum in Japan. In association with this reduction in genetic diversity was a significantly higher inter-colony fusion rate between randomly paired New Zealand D. vexillum colonies (80%, standard deviation ±18%) when compared with colonies found in Japan (27%, standard deviation ±15%). The results of this study add to growing evidence that for colonial organisms reductions in population level genetic diversity may alter colony interaction dynamics and enhance the invasive potential of newly colonizing species.     

Genotypic variability following fusion in the invasive colonial tunicate Didemnum vexillum

Fusion to form a chimera has been documented in many marine invertebrate taxa, including poriferans, cnidarians, bryozoans, and colonial ascidians. Allogenic interactions in chimeric ascidian colonies vary widely across taxonomic groups but are poorly characterized in the invasive colonial ascidian Didemnum vexillum. The moderate level of discrimination expressed in the fusion–rejection response of D. vexillum suggests that there is some integration of cells beyond the fusion line in a chimeric colony. We tracked the shifts in representation of microsatellite alleles between fused colonies of D. vexillum to elucidate the extent of genotypic integration in fused colonies and the patterns of changes to the genotypic composition of colonies immediately following chimera formation. By genotyping colonies before and after fusion, we found that allogeneic fusion in D. vexillum may lead to genotypic changes beyond the visible fusion line. Alleles from one colony were found in multiple tissue samples in the chimera 7–10 days after fusion had occurred. In some instances, alleles that were in a single colony prior to fusion were lost following fusion. We observed multiple patterns of allelic change, including both the unidirectional transfer and reciprocal exchange of alleles between fused colonies. Our findings suggest that tissue or cells are exchanged following allogeneic fusion between colonies of D. vexillum and that the genotypic composition of chimeric colonies may be fluid.     

Tunic cell populations during fusion events in the colonial ascidian Didemnum vexillum (Tunicata)

We documented changes in the abundance and distribution patterns of tunic cells involved in the allorecognition response of the colonial aplousobranch Didemnum vexillum, whose zooids do not share a common vascular system. A histological examination of the fusion zone of isogeneic (CIAs) and allogeneic (CAAs) fused colony assays revealed that tunic cuticles were rapidly regenerated. The underlying tunic matrix fused readily in all assays and controls. We identified four different types of tunic cells. Phagocytic cells represented the most abundant cell type in allogeneic fusions, followed by morula cells. These cells were more abundant at the immediate fusion junction than at 120 μm or 240 μm from the junction, most likely because they mediate the allorecognition reaction. Elongated filopodial cells also were present, although only at very low abundances, and a layer of bladder cells was located immediately below the cuticle. Our results provide quantitative evidence for the involvement of tunic cells in the allorecognition response of a highly invasive ascidian.     

Predation Limits Spread of Didemnum vexillum into Natural Habitats from Refuges on Anthropogenic Structures

Non-indigenous species can dominate fouling assemblages on artificial structures in marine environments; however, the extent to which infected structures act as reservoirs for subsequent spread to natural habitats is poorly understood. Didemnum vexillum is one of few colonial ascidian species that is widely reported to be highly invasive in natural ecosystems, but which in New Zealand proliferates only on suspended structures. Experimental work revealed that D. vexillum established equally well on suspended artificial and natural substrata, and was able to overgrow suspended settlement plates that were completely covered in other cosmopolitan fouling species. Fragmentation led to a level of D. vexillum cover that was significantly greater than was achieved as a result of ambient larval recruitment. The species failed to establish following fragment transplants onto seabed cobbles and into beds of macroalgae. The establishment success of D. vexillum was greatest in summer compared with autumn, and on the underside of experimental settlement plates that were suspended off the seabed to avoid benthic predators. Where benthic predation pressure was reduced by caging, D. vexillum establishment success was broadly comparable to suspended treatments; by contrast, the species did not establish on the face-up aspect of uncaged plates. This study provides compelling evidence that benthic predation was a key mechanism that prevented D. vexillum’s establishment in the cobble habitats of the study region. The widespread occurrence of D. vexillum on suspended anthropogenic structures is consistent with evidence for other sessile invertebrates that such habitats provide a refuge from benthic predation. For invasive species generally, anthropogenic structures are likely to be most important as propagule reservoirs for spread to natural habitats in situations where predation and other mechanisms do not limit their subsequent proliferation.     

Genetic diversity and relatedness in aquaculture and marina populations of the invasive tunicate Didemnum vexillum in the British Isles

Introductions of invasive, non-native species in the marine environment are increasing as human activity within coastal areas rises. Genetic datasets are useful tools to identify source populations, track routes of invasions, and illuminate the role of genetic variation in the establishment and subsequent spread of novel introductions. Here, a microsatellite dataset is used to estimate the genetic diversity and population structure of 7 introduced Didemnum vexillum populations in Britain and Ireland, 4 of which are associated with aquaculture and 3 with marinas. Genetic differentiation observed between these populations indicates human-mediated transport as the main mechanism underlying the population structure of D. vexillum in Britain and Ireland. In addition to elucidating patterns of population structure we found that aquaculture sites showed significantly higher genetic diversity (measured as allelic richness) in comparison to the marina sites. We discuss these findings in relation to the history of each invasion, the complex life history of D. vexillum, and available evidence of the relative invasiveness of these populations. Our results show numerous interesting patterns which highlight further research avenues to elucidate the complex factors underlying the global spread of this successful invader.

     

New GlcNAc/GalNAc-specific lectin from the ascidian Didemnum ternatanum

Previously we isolated GlcNAc-specific lectin (DTL) from the ascidian Didemnum ternatanum by affinity chromatography on cross-linked ovalbumin. Here we report the purification and characterization of new D-GlcNAc/D-GalNAc-specific lectin DTL-A from the same ascidian. This lectin was isolated from non-bound cross-linked ovalbumin fraction and further was purified by gel filtration on Sepharose CL-4B, affinity chromatography on GlcNAc-agarose and gel filtration on Superdex 200. SDS-polyacrylamide gel electrophoresis and gel filtration of purified lectin on Sepharose CL-4B indicates that it exists as large aggregates in the native state. Investigations of the carbohydrate specificity of DTL-A by enzyme-linked lectin assay suggest the multi-specificity of this lectin. DTL-A binds BSM, asialo-BSM as well as heparin and dextran sulfate. The binding of DTL-A to BSM was inhibited by monosaccharides D-GlcNAc and D-GalNAc, their alpha- but not beta-anomers. Among polysaccharides and glycoconjugates, DTL-A binding to BSM was effectively inhibited by BSM, asialo-BSM, pronase-treated BSM and synthetic alpha-D-GalNAc-PAA. Fetuin and asialofetuin showed a much lower inhibitory potency, heparin and dextran sulfate were noninhibitory. On the other hand, DTL-A binding to heparin was effectively inhibited by dextran sulfate, fucoidan, whereas BSM showed insignificantly inhibitory effect. DTL-A binding to heparin was not inhibited by D-GlcNAc and D-GalNAc.     

UV protection in the photosymbiotic ascidian Didemnum molle inhabiting different depths

Didemnum molle is a colonial ascidian that harbors the prokaryotic photosymbiont Prochloron in its cloacal cavity. Colonies occur over a relatively wide bathymetric range (approximately 0-30 m), and colony color is widely variable, partly depending on depth. Colonies in shallow sites are bright white, with densely distributed spicules, and often with brown or dark gray pigmentation, while colonies in deeper sites are less pigmented, with sparsely distributed spicules. Didemnum molle colonies contain mycosporine-like amino acids (MAAs) as UV-absorbing substances. These include mycosporine-glycine, shinorine, and porphyra-334. Among colonies from 5-, 10-, 15-, and 20-m depths, the concentration of total MAAs was significantly high at 10 m and low at 20 m. Colonies at 10 m need to maintain low spicule densities to have enough photosynthetically active radiation (PAR) to maintain the photosymbionts, and they probably concentrate MAAs to block UV radiation without attenuating PAR. Because high levels of PAR cause photoinhibition of photosynthesis, spicules and pigment cells would be more effective for photoprotection in shallow water. Colonies of D. molle may adjust the light conditions for photosymbionts by combining MAAs, spicules, and pigment cells in varying amounts.     

Contents of ultraviolet-absorbing substances in two color morphs of the photosymbiotic ascidian Didemnum molle

The contents of mycosporine-like amino acids (MAAs) were compared in the two color morphs (dark-gray and brown colonies) of the tropical ascidian Didemnum molle (Herdman, 1886), which harbors the photosymbiotic prokaryote Prochloron. The colonies of each color morph were exclusively distributed in shallow reef lagoons at the different sites. Spectroscopic and chromatographic analyses showed that the Prochloron cell density and MAA concentration in the dark-gray colonies were an estimated 1.4 and 2.4 times higher, respectively, than in the brown colonies. The significant difference in MAA contents between the color morphs was primarily due to the difference in shinorine contents (p < 0.01, Mann–Whitney U-test). The high concentration of MAAs in the dark-gray colonies may provide better conditions for Prochloron cells, compared to the brown colonies with lower MAA concentrations.     

Range expansion and potential dispersal mechanisms of the exotic cladoceran Daphnia lumholtzi

. Recently the exotic cladoceran Daphnia lumholtzi has invadedfreshwater systems throughout the southern and midwestern UnitedStates. We conducted regional surveys of eastern Kansas reservoirsto document the range expansion of D.lumholtzi. Daphnia lumholtziwas found in five of 35 reservoirs sampled in 1994, and 11 ofthe 35 reservoirs when re-sampled in 1997. In addition, we sampled40 small ponds inaccessible to recreational boats, within thewatershed of an invaded reservoir. We did not find D.lumholtziin any of these ponds, suggesting that non-human dispersal mechanismsplay an insignificant role in the range expansion of D.lumholtzithroughout the United States. Further experimentation, however,is needed to determine if the absence of D.lumholtzi from thesesmall ponds is due to insufficient dispersal mechanisms or theinability of this invader to successfully colonize followingarrival. Daphnia lumholtzi has broad limnological tolerances.However, invaded reservoirs tend to be larger in area, havehigher Secchi disk depths, and lower total phosphorus, totalnitrogen and chlorophyll a levels relative to non-invaded reservoirs.Analyses of pre-invasion zooplankton communities indicate thatD.lumholtzi may be invading reservoirs in which native Daphniaspecies are rare. While the long-term effects of the invasionof D.lumholtzi are unknown, it has the potential to dominatelate summer zooplankton communities in eastern Kansas reservoirs.Therefore, we need to continue to survey and monitor invadedreservoirs to document the range expansion of D.lumholtzi anddetermine the long-term implications of the introduction ofthis invader.     

Real‐time PCR detection of Didemnum perlucidum (Monniot, 1983) and Didemnum vexillum (Kott, 2002) in an applied routine marine biosecurity context

Prevention and early detection are well recognized as the best strategies for minimizing the risks posed by nonindigenous species (NIS) that have the potential to become marine pests. Central to this is the ability to rapidly and accurately identify the presence of NIS, often from complex environmental samples like biofouling and ballast water. Molecular tools have been increasingly applied to assist with the identification of NIS and can prove particularly useful for taxonomically difficult groups like ascidians. In this study, we have developed real‐time PCR assays suited to the specific identification of the ascidians Didemnum perlucidum and Didemnum vexillum. Despite being recognized as important global pests, this is the first time specific molecular detection methods have been developed that can support the early identification and detection of these species from a broad range of environmental sample types. These fast, robust and high‐throughput assays represent powerful tools for routine marine biosecurity surveillance, as detection and confirmation of the early presence of species could assist in the timely establishment of emergency responses and control strategies. This study applied the developed assays to confirm the ability to detect Didemnid eDNA in water samples. While previous work has focused on detection of marine larvae from water samples, the development of real‐time PCR assays specifically aimed at detecting eDNA of sessile invertebrate species in the marine environment represents a world first and a significant step forwards in applied marine biosecurity surveillance. Demonstrated success in the detection of D. perlucidum eDNA from water samples at sites where it could not be visually identified suggests value in incorporating such assays into biosecurity survey designs targeting Didemnid species.     

Dispersal patterns, dispersal mechanisms, and invasion wave speeds for invasive thistles

Understanding and predicting population spread rates is an important problem in basic and applied ecology. In this article, we link estimates of invasion wave speeds to species traits and environmental conditions. We present detailed field studies of wind dispersal and compare nonparametric (i.e., data-based) and mechanistic (fluid dynamics model-based) dispersal kernel and spread rate estimates for two important invasive weeds, Carduus nutans and Carduus acanthoides. A high-effort trapping design revealed highly leptokurtic dispersal distributions, with seeds caught up to 96 m from the source, far further than mean dispersal distances (approx. 2 m). Nonparametric wave speed estimates are highly sensitive to sampling effort. Mechanistic estimates are insensitive to sampling because they are obtained from independent data and more useful because they are based on the dispersal mechanism. Over a wide range of realistic conditions, mechanistic spread rate estimates were most sensitive to high winds and low seed settling velocities. The combination of integrodifference equations and mechanistic dispersal models is a powerful tool for estimating invasion spread rates and for linking these estimates to characteristics of the species and the environment.     

Comparing the potential for dispersal via waterbirds of a native and an invasive brine shrimp

1. Migratory waterbirds are likely to have a major role in the spread of many exotic aquatic invertebrates by passive dispersal. However, in the field, this has so far only been confirmed in the case of the American brine shrimp Artemia franciscana, which is spreading quickly around the Mediterranean region. 2. We compared experimentally the capacity of A. franciscana and the native brine shrimp Artemia parthenogenetica to disperse via migratory shorebirds. After Artemia resting eggs (cysts) were fed to Redshank Tringa totanus and Dunlin Calidris alpina, we compared the proportion that survived gut passage, their hatchability and their retention time within the gut. We also tested the ability of cysts to stick to the feathers of Black‐tailed Godwit Limosa limosa. 3. The proportion of ingested cysts retrieved from faeces was the same for each Artemia species (8%), and there were no significant differences in retention time (mean 1.2 h and maximum 10 h for A. parthenogenetica, 1.4 and 12 h for A. franciscana) or hatchability (11% versus 14%). The two shorebird species showed similar retention times and retrieval rates, but cysts recovered from Dunlin had a significantly higher hatchability. Only one of the 1000 A. parthenogenetica cysts and three of the 1000 A. franciscana cysts stuck to feathers. 4. These results indicate that both non‐native and native brine shrimps have a similar high capacity for endozoochory via birds, and that the invasiveness of A. franciscana is probably explained by its competitive superiority owing to high fecundity and release from cestode parasitism. Owing to their different migratory behaviour, Redshank and Dunlin are likely to have different roles as brine shrimp vectors. Brine shrimps provide a suitable model for understanding the role of birds in the dispersal of exotic aquatic invertebrates.     

Natural and anthropogenic dispersal mechanisms in the marine environment: a study using cheilostome Bryozoa

The global geographic ranges occupied by 197 species of cheilostomate Bryozoa found in British waters were obtained by a literature survey. Morphological grade, larval mode, environmental tolerance, species abundance and the ability to raft and to foul shipping were all investigated as traits potentially able to affect the geographic ranges of these bryozoan species. When considered independently all variables except larval mode had a significant correlation with the geographic range occupied by a species. However, when controlling for the potentially confounding effects of the other covariates, only the ability to foul or raft and species abundance had a significant effect on median geographic range and only fouling and abundance had a significant effect over global ranges. The strength of the association between fouling ability and range suggests that transport upon the hulls of ships is a very important dispersal mechanism for bryozoans, as it is thought to be also for various other marine taxa. Potential long-term (evolutionary) consequences of increased ranges brought about by anthropogenic mechanisms are discussed.     

Spatial competition induces the mobilization of morula cells in the colonial ascidian Didemnum perlucidum (Tunicata: Didemnidae)

Abstract. The effects of spatial competition among colonial marine organisms are often evident in the contact zones between colonies. These effects are especially pronounced when the interaction results in overgrowth or necrosis of one of the competitors. Ascidians, one of the dominant taxonomic groups in subtidal sessile communities, have specialized morula cells that provide a defense against microbial infections. Injuries resulting from interspecific competitive interactions might also act as a stimulus for this defensive mechanism. Therefore, we expected to see the recruitment of morula cells in tissues near competitor contact zones. To test the hypothesis that spatial competition elicits this immune response, we placed colonies of the ascidian Didemnum perlucidum from southeastern Brazil in four different types of competitive situations: (1) overgrowth of the competitor, (2) stand‐off interactions, (3) overgrowth by the competitor, and (4) free of competitors. Our results indicate that competitive interactions increase the population of morula cells in contact zones, as more cells were observed in interactions that resulted in the overgrowth of individuals of D. perlucidum, and fewer cells were observed in colonies that were free of competitors. We identified the defensive function of the morula cells by showing the presence of the enzyme phenoloxidase within its vacuoles. Phenoloxidase is a widespread enzyme among animals and plants, and is frequently used in defense by synthesizing toxic quinones from polyphenol substrates. This is the first study to document the presence of morula cells in didemnid ascidians and the mobilization of these cells by spatial competition by heterospecifics, and one of the first studies to identify phenoloxidase activity in morula cells.     

Spread pathways of the invasive weed Parthenium hysterophorus L.: The potential for water dispersal

Parthenium hysterophorus L. (Asteraceae) utilises multiple mechanisms to facilitate its dispersal. It has been speculated that the cypsela, the propagule of this species, can be dispersed by water under varying environmental conditions. Four experiments were conducted to test this hypothesis, using simulated shaking and immersion to test floating ability and viability of the propagule in water. The influence of the acidity of the immersion medium on cypsela viability was also examined. Our results revealed that the freshly harvested cypselae could float on river water for at least 20 days, although around 80% sank within a week if moderate or severe turbulence was applied. Sinkage was observed to be more rapid in naked seeds (within a day) than in cypsela (within a week). On still water surfaces, germination occurred within a week but extended to 1.5 weeks under turbulent conditions due to sinkage. In river water, initial germination of floating cypselae was greater (70%) under illuminated conditions as compared to dark conditions (20%). The viability of immersed cypselae was found to remain high in distilled water for 45 days, when immersion was in cool conditions (10 or 15°C). However, in moderate (20 and 24°C) or warm (25 and 30°C) conditions, the rate of viability loss increased, and at 34°C, around 50% of the cypselae died after 20 days of immersion. Similar trends for cypselae longevity were observed in studies using river and pond water; viability loss was faster, especially in pond water. In summary, a proportion of cypselae will float in turbulent water and could be carried significant distances in river systems. Immersed cypselae can remain viable for weeks and can germinate on contact with soil. Water bodies or floods are therefore considered as important pathways in parthenium weed dispersal; hence, post‐flood monitoring is strongly recommended to minimise its spread.     

Natural products with anti-aging potential: Affected targets and molecular mechanisms

In recent years, there has been a great deal of attention toward the molecular machinery relevant to age-related progression controlled through the external intervention of polyphenols- an epigenetic-modulating diet. Natural products modulate cellular longevity through histone post-translational modification and can also induce the upregulation of autophagy, thus reducing the level of acetyl coenzyme A (AcCoA). In addition, the effect of caloric restriction (CR) on cancer-related chronic inflammation is of great significance in aging. In line with this, SIRT1 protein levels are expanded in response to calorie restriction mimetics (CRM), in this way acting as autophagy inducers relevant to cancer prevention.     

Genetic analysis across different spatial scales reveals multiple dispersal mechanisms for the invasive hydrozoan Cordylophora in the Great Lakes

Discerning patterns of post‐establishment spread by invasive species is critically important for the design of effective management strategies and the development of appropriate theoretical models predicting spatial expansion of introduced populations. The globally invasive colonial hydrozoan Cordylophora produces propagules both sexually and vegetatively and is associated with multiple potential dispersal mechanisms, making it a promising system to investigate complex patterns of population structure generated throughout the course of rapid range expansion. Here, we explore genetic patterns associated with the spread of this taxon within the North American Great Lakes basin. We collected intensively from eight harbours in the Chicago area in order to conduct detailed investigation of local population expansion. In addition, we collected from Lakes Michigan, Erie, and Ontario, as well as Lake Cayuga in the Finger Lakes of upstate New York in order to assess genetic structure on a regional scale. Based on data from eight highly polymorphic microsatellite loci we examined the spatial extent of clonal genotypes, assessed levels of neutral genetic diversity, and explored patterns of migration and dispersal at multiple spatial scales through assessment of population level genetic differentiation (pairwise F_ST and factorial correspondence analysis), Bayesian inference of population structure, and assignment tests on individual genotypes. Results of these analyses indicate that Cordylophora populations in this region spread predominantly through sexually produced propagules, and that while limited natural larval dispersal can drive expansion locally, regional expansion likely relies on anthropogenic dispersal vectors.     

Short- and long-range dispersal of medfly, Ceratitis capitata (Dipt., Tephritidae), and its invasive potential

Abstract:  Data were obtained from mark recapture trials pertaining to the dispersal of medfly, Ceratitis capitata (Dipt., Tephritidae), over both short (10–160 m) and very long distances (0.5–9.5 km) within the surveillance trapping array in Adelaide, Australia. They could be related to previously reported data sets by expressing the capture rates of each set in common terms that corrected for differences in recapture rate resulting from type of trap, season or climate. The mean capture rate at each distance from the point of release in each data set was expressed as a percentage of the real or inferred rate of that set at a distance of 100 m. The resulting distribution of dispersal distances conformed to both an inverse power model and a modified Cauchy model regardless of whether the present and previous data were combined or not. The modified Cauchy model inferred that the median distance flown was extremely short and 90% of flies displaced only 400–700 m despite the fact that a consistent trend in declining catch rates was obtained up to 9.5 km. The spread of invading propagules in quarantined zones in the first generation is likely to be limited by a decline to non-viable density within 1 km or less of the incursion point and the spread of larger infestations could be limited by the longevity of the dispersers. The results also have significance to the ability of surveillance trapping arrays to detect infestations and also to methods of distributing insects for the 'sterile insect technique'.