Post-exchange zooplankton in ballast water of ships entering the San Francisco Estuary

The San Francisco Estuary in California (CA), USA, has beenheavily altered by invasions of nonnative zooplankton and benthicorganisms, presumably by the discharge of ships’ ballastwater. Since 2000, ships entering CA have been required to exchangeballast water with oceanic water during the voyage to decreasethe number of organisms discharged into the Estuary that hadpreviously been taken aboard at foreign ports. We examined abundanceof zooplankton in ballast water of 18 container ships and 48bulk carriers. Asia dominated the sources of ballast water,which contained multiple nonnative zooplankton including speciesthat have invaded and since become common residents in the Estuary.The abundance of zooplankton was significantly lower in ballastwater that had been emptied and refilled with oceanic waterthan those that had continuously been flushed with oceanic water(about three times the volume of ballast water), suggestingthat empty–refill is more effective in removing exoticzooplankton.     

Population attenuation in zooplankton communities during transoceanic transfer in ballast water

Successful biological invasion requires introduction of a viable population of a nonindigenous species (NIS). Rarely have ecologists assessed changes in populations while entrained in invasion pathways. Here, we investigate how zooplankton communities resident in ballast water change during transoceanic voyages. We used next‐generation sequencing technology to sequence a nuclear small subunit ribosomal DNA fragment of zooplankton from ballast water during initial, middle, and final segments as a vessel transited between Canada and Brazil. Operational taxonomic unit (OTU) diversity decreased as voyage duration increased, indicating loss of community‐based genetic diversity and development of bottlenecks for zooplankton taxa prior to discharge of ballast water. On average, we observed 47, 26, and 24 OTUs in initial, middle, and final samples, respectively. Moreover, a comparison of genetic diversity within taxa indicated likely attenuation of OTUs in final relative to initial samples. Abundance of the most common taxa (copepods) declined in all final relative to initial samples. Some taxa (e.g., Copepoda) were represented by a high number of OTUs throughout the voyage, and thus had a high level of intraspecific genetic variation. It is not clear whether genotypes that were most successful in surviving transit in ballast water will be the most successful upon introduction to novel environments. This study highlights that population bottlenecks may be common prior to introduction of NIS to new ecosystems.     

Species Richness and Invasion Vectors: Sampling Techniques and Biases

During a European Union Concerted Action study on species introductions, an intercalibration workshop on ship ballast water sampling techniques considered various phytoplankton and zooplankton sampling methods. For the first time, all the techniques presently in use worldwide were compared using a plankton tower as a model ballast tank spiked with the brine shrimp and oyster larvae while phytoplankton samples were taken simultaneously in the field (Helgoland Harbour, Germany). Three cone-shaped and 11 non-cone shaped plankton nets of different sizes and designs were employed. Net lengths varied from 50 to 300 cm, diameters 9.7–50 cm, and mesh sizes 10–100 μm. Three pumps, a Ruttner sampler, and a bucket previously used in ballast water sampling studies were also compared. This first assessment indicates that for sampling ballast water a wide range of techniques may be needed. Each method showed different results in efficiency and it is unlikely that any of the methods will sample all taxa. Although several methods proved to be valid elements of a hypothetical `tool box' of effective ship sampling techniques. The Ruttner water sampler and the pump P30 provide suitable means for the quantitative phytoplankton sampling, whereas other pumps prevailed during the qualitative trial. Pump P15 and cone-shaped nets were the best methods used for quantitative zooplankton sampling. It is recommended that a further exercise involving a wider range of taxa be examined in a larger series of mesocosms in conjunction with promising treatment measures for managing ballast water. This revised version was published online in July 2006 with corrections to the Cover Date.     

Survival of tropical ballast water organisms during a cruise from the Indian Ocean to the North Sea

In an assessment of non-indigenous species transported by internationalship traffic to German waters, commissioned by the German FederalEnvironmental Agency, the survival of tropical plankton organismsin ballast water was studied by accompanying a container vesselon its 23-day voyage from Singapore to Bremerhaven in Germany.Two tanks, one filled off Singapore and the other off Colombo,Sri Lanka, were monitored for their phyto- and zooplankton contentby daily sampling. As already reported in previous studies,species abundance and diversity, especially of zooplankton,decreased sharply during the first days, and only a few specimenssurvived the whole cruise. The contents of the Colombo tank,however, changed dramatically during the last week. The harpacticoidcopepod, Tisbe graciloides, increased its abundance by a factorof 100 from 0.1 to 10ind. l^–1 within a few days. Thisis the first time that a ballast water organism has been foundto multiply at such a high rate. Opportunistic species suchas Tisbe are apparently able to thrive and propagate in ballastwater tanks under certain conditions. Ballast water tanks maythus serve as incubators for certain species depending on theircharacteristics.     

A biological survey of ballast water in container ships entering Hong Kong

The role of ballast water in the introduction of exotic species has recently received extensive attention. The aim of this study is to assess the importance of ballast water discharge as a vector for the introduction of exotic species into Hong Kong waters. Twelve ballast water samples were collected from 5 container ships entering Hong Kong between June1994 and October 1995. The ballast water originated from ports on both sides of the Pacific Ocean. At least 81 species from 8 animal phyla and 5 protist phyla were found. Most of the major marine taxonomic groups were represented and many planktonic larval stages were included. Species richness in the ballast tanks decreased with the age of ballast water. Copepoda was the most diverse and abundant taxonomic group. The density of calanoid and cyclopoid copepods decreased with the age of ballast water, but that of harpacticoid copepods did not change significantly with time. Bivalve, crustacean, polychaete and ascidian larvae from ballast water samples were observed to settle in laboratory culture tanks. The mussel Mytilopsis sallei which was introduced to Hong Kong in 1980, was one of the bivalves that settled readily. Results of this study indicate that ballast water can be a major source for the introduction of exotic species to Hong Kong waters. Regulatory guidelines on the discharge of ballast water should be established. This revised version was published online in August 2006 with corrections to the Cover Date.     

Diatom survivorship in ballast water during trans-Pacific crossings

Ship ballast water is believed to be responsible for global dispersal of alien biota; mid-ocean ballast water exchange is most commonly used to mitigate this process. Diatoms are among the most abundant biotic-component in ballast water, yet their invasive biology is poorly understood. To test effectiveness of MOE we examined diatom species composition and cell density in two sets of samples. First, we examined samples collected daily during one 24 days long trans-Pacific crossing in tanks with and without ballast exchanged. Second, we used samples from 23 trans oceanic vessels arriving at Vancouver harbour where diatoms were collected on arrival. Up to 86,429 live diatom cells/l were found in the tanks, ~50% of the samples share up to eight species consistently present. Cell densities and species richness declined over time and with replacement of coastal ballast water by mid-oceanic water. In both data sets diatoms survive in the tanks for as long as 33 days despite ballast exchange.     

Flow cytometry, microscopy, and DNA analysis as complementary phytoplankton screening methods in ballast water treatment studies

Ballast water is the main vector for marine invasions. To minimize the spread of invasive species, the International Maritime Organization (IMO) has adopted the Ballast Water Management Convention which requires the installation of shipboard ballast water treatment systems (BWTS). During BWTS tests, the phytoplankton abundance and species composition were followed after treatment with both filtration and ultraviolet radiation. Although the installation fulfilled the IMO criteria after a 5-day holding time in a model ballast tank, the ultimate effectiveness of the treatment was further tested in long-term (20 days) incubation experiments under optimal phytoplankton growth conditions. Application of flow cytometry, microscopy, and DNA sequencing to these incubation samples gave an indication of the phytoplankton species that might be introduced by ballast water discharge—despite treatment. Phytoplankton was reliably quantified using flow cytometry, while fast identification was best done using microscopy. Some groups that contained potentially toxic species could not be identified at species level using microscopy; for these species, identification using genetic techniques was necessary. It is concluded that if long-term incubation experiments are used as an additional tool in testing BWTS effectiveness, a combination of phytoplankton screening methods can be applied depending on the detail of information that is required.     

Advective effects of a reservoir flood on zooplankton abundance and dispersion

SUMMARY. 1. The diel vertical distribution and abundance of zooplankton in the Red River arm of Lake Texoma, Oklahoma-Texas, U.S.A., were examined between 25 January and 26 June 1982.
2. During periods of low river inflow, zooplankton vertical patchiness was related to wind stress. Zooplankton did not maintain distinct vertical patches under strong mixing.
3. In mid May heavy rainfall in the drainage basin increased discharge from the Red River and reduced water retention time in the reservoir. The Red River water mass entered the lake as a turbid, surface overflow plume. During this period of flooding most zooplankton populations declined and the distributions of remaining individuals deepened. Greatest population losses occurred between 0 and 7m, depths where most zooplankton had been concentrated prior to the flood.
4. A model of population dynamics based on depth specific rates of water exchange between the lake and the Red River, estimated from conductivity changes through time, accounted for the general pattern of change in zooplankton abundance and vertical dispersion.     

The efficiency of regional ballast water exchange: Changes in phytoplankton abundance and diversity

Ballast water was sampled on 12 occasions before and after an exchange process carried out in regional seas in order to assess the efficiency of this type of ballast water management at reducing the abundance and diversity of phytoplankton. Although there was an overall reduction in the average abundance and number of taxa after exchange this was not consistent between tanks and voyages. On some occasions there were changes in species composition after exchange and, in some cases, there were increases in potentially harmful species after the exchange process. Factors such as the depth of the water during the exchange process, the season and the method of exchange influenced the efficacy of the exchange process. The variability in the results after exchange mean that this is unlikely to be a ballast water management method that would give consistent results and careful consideration would have to be given to the suitability of using this method in regional seas as a means of reducing the risk of introducing non-native species.     

Efficacy of ‘saltwater flushing’ in protecting the Great Lakes from biological invasions by invertebrate eggs in ships’ ballast sediment

1. Mid‐ocean exchange and saltwater flushing were implemented as management practices to reduce the likelihood of new biological invasions in the Laurentian Great Lakes associated with ships’ ballast water and sediments. Despite this, there has been no formal assessment of the efficacy of these procedures. Here, we conduct a comparative analysis of community composition of dormant taxa transported by ballast sediment before and after regulations came into effect in 2006. 2. Ballast sediment samples were collected from 17 ships during the post‐regulation interval of 2007 and 2008. Invertebrate eggs were counted, hatched and species identified in the laboratory. Results were compared to similar samples collected from 39 ships between 2000 and 2002, prior to implementation of saltwater flushing regulations. 3. The estimated amount of residual ballast sediment transported by vessels was significantly lower during the post‐regulation period, ranging from <1 to 45 tonnes per ship, with an average of 5 tonnes. Mean density and number of dormant viable eggs per ship declined 91 and 81%, respectively. 4. Community composition also changed through time, with Rotifera accounting for 78% of taxa transported prior to regulation, whereas Cladocera and Copepoda each accounted for 38% of abundance post‐regulation. Although the number of non‐indigenous species (NIS) declined 73% per ship after 2006, the reduction was not statistically significant; however, the number of freshwater NIS – which pose the greatest risk of invasion for the Great Lakes – was significantly lowered. 5. Our comparative analysis suggests that ballast management regulations enacted in 2006 markedly reduced the probability of introduction of NIS via dormant eggs carried in ballast sediments.     

Transport of diatom and dinoflagellate resting spores in ships' ballast water: implications for plankton biogeography and aquaculture

Diatom and dinoflagellate species that are not endemic to aregion can be inadvertently introduced when their resistantresting stages are discharged with the ballast-tank waters andsediments of bulk cargo vessels. A survey of 343 cargo vesselsentering 18 Australian ports showed that 65% of ships were carryingsignificant amounts of sediment on the bottom of their ballasttanks. All of these samples contained diatoms, including speciesthat are not endemic to Australian waters. Diatom resting spores,especially of Chaetoceros, were also detected. Dinoflagellateresting spores (cysts) were present in 50% of the sediment samples.Of the 53 cyst species identified, 20 (including Diplopelta,Diplopsalopsis, Gonyaulax, Polykrikos, Protoperidinium, Scrippsiellaand Zygabikodinium spp.) were successfully germinated to produceviable cultures. Such diversity of diatom and dinoflagellatespecies in ships' ballast water suggests that the apparent cosmopolitanismof many coastal phytoplankton species may be due partly to theglobal transport of seawater ballast. Of considerable concernwas the detection in 16 ships of cysts of the toxic dinoflagellatesAlexandrium catenella, Alexandrium tamarense and Gymnodiniumcatenatum. One single ballast tank was estimated to contain>300 million viable A.tamarense cysts, some of which weresuccessfully germinated in the laboratory to produce toxic cultures.These toxic dinoflagellate species, which can contaminate shellfishwith paralytic shellfish poisons, pose a serious threat to humanhealth and the aquaculture industry. Ballast-water quarantinemeasures recently introduced in Australia are discussed. Mid-oceanexchange of ballast water is only partially effective in removingdinoflagellate cysts which have settled to the bottom of ballasttanks. The present work indicates that the most effective measureto prevent the spreading of toxic dinoflagellate cysts via ships'ballast water would be to avoid taking on ballast water duringdinoflagellate blooms in the water column of the world's ports.     

Studies on Plover Cove Reservoir, Hong Kong

The zooplankton fauna of the Plover Cove Reservoir has been investigated using two sampling series. The first series involving a 3-year investigation using 6-ft vertical hauls at three stations in the reservoir, revealed the presence of two copepod and four cladoceran species with little variation in terms of seasonal peak occurrence and abundance in any of the stations and so no evidence of the horizontal distribution patterns referred to by other authors. In all three stations Diaptomus gracilis was the dominant copepod species and Diaphanosoma leuchtenbergianum the dominant cladoceran species. The second series, involving a 13-month investigation of vertical sets of water samples collected both day and night at 4-h intervals for 24-h periods, revealed that in terms of seasonal vertical distribution, all six species occurred at all depths during the survey, but with a tendency to be concentrated in the surface waters. In terms of diurnal vertical distribution, differences in the migration pattern occurred from season to season in the case of Diaptomus gracilis. Thus, during spring, summer and autumn, concentrations occurred at the surface at 10.00 and 22.00 hours followed by migration to greater depth, whereas in winter, surface accumulations persisted even into the daylight hours. However, both Cyclops hyalinus and Diaphanosoma leuchtenbergianum showed surface accumulations at night and a movement away from the surface by day during all seasons. Investigations of the seasonal variation in environmental parameters conducted during both sampling series revealed that, as in the case of similar surveys, water temperature was the major controlling factor governing the seasonal variation in composition of the zooplankton fauna as well as its seasonal vertical distribution since in the case of Diaptomus gracilis and Ceriodaphnia reticulata low water temperatures corresponded with peak seasonal occurrence whereas in the case of the other species high water temperatures corresponded with peak seasonal occurrence, and since Diaptomus gracilis and Diaphanosoma leuchtenbergianum showed maximum concentration near the surface when surface water temperatures were high, whereas the remaining species showed their maximum concentration near the surface when lower water temperatures prevailed. It would also appear, however, that seasonal changes in zooplankton densities closely followed algal densities since Diaptomus gracilis, Diaphanosoma leuchtenbergianum and Bosmina longirostris had their maxima in late spring and Cyclops hyalinus in late summer while algal densities in this reservoir were highest in early spring and early summer. With regard to the diurnal vertical distribution of the zooplankton, light appeared to be the only factor alternating by an amount likely to be correlated with such diurnal vertical changes in zooplankton density.     

Habitat selection and diel distribution of the crustacean zooplankton from a shallow Mediterranean lake during the turbid and clear water phases

1. The fish fauna of many shallow Mediterranean Lakes is dominated by small‐bodied exotic omnivores, with potential implications for fish–zooplankton interactions still largely unknown. Here we studied diel variation in the vertical and horizontal distribution of the crustacean plankton in Lake Vela, a shallow polymictic and eutrophic lake. Diel sampling was carried out on three consecutive days along a horizontal transect, including an open‐water station and a macrophyte (Nymphaea alba) bed. Since transparency is a key determinant of the predation risk posed by fish, the zooplankton sampling campaigns were conducted in both the turbid (autumn) and clear water (spring) phases. 2. In the turbid phase, most taxa were homogeneously distributed along the vertical and horizontal axes in the three consecutive days. The only exception was for copepod nauplii, which showed vertical heterogeneity, possibly as a response to invertebrate predators. 3. In the clear water phase, most zooplankton taxa displayed habitat selection. Vertically, the general response consisted of a daily vertical migration (DVM), despite the limited depth (1.6 m). Horizontally, zooplankters showed an overall preference for the pelagic zone, independent of the time of the day. Such evidence is contrary to the postulated role of macrophytes as an anti‐predator refuge for the zooplankton. 4. These vertical (DVM) and horizontal (macrophyte‐avoidance) patterns were particularly conspicuous for large Daphnia, suggesting that predation risk from size‐selective predators (fish) was the main factor behind the spatial heterogeneity of zooplankton in the spring. Thus, the difference in the zooplankton spatial distribution pattern and habitat selection among seasons (turbid and clear water phases) seems to be mediated the predation risk from fish, which is directly related to water transparency. 5. The zooplankton in Lake Vela have anti‐predator behaviour that minimises predation from fish. We hypothesise that, due to the distinct fish community of shallow Mediterranean lakes, aquatic macrophytes may not provide adequate refuge to zooplankters, as seen in northern temperate lakes.     

Molecular detection of native and invasive marine invertebrate larvae present in ballast and open water environmental samples collected in Puget Sound

Non-native marine species have been and continue to be introduced into Puget Sound via several vectors including ship's ballast water. Some non-native species become invasive and negatively impact native species or near shore habitats. We present a new methodology for the development and testing of taxon specific PCR primers designed to assess environmental samples of ocean water for the presence of native and non-native bivalves, crustaceans and algae. The intergenic spacer regions (IGS; ITS1, ITS2 and 5.8S) of the ribosomal DNA were sequenced for adult samples of each taxon studied. We used these data along with those available in Genbank to design taxon and group specific primers and tested their stringency against artificial populations of plasmid constructs containing the entire IGS region for each of the 25 taxa in our study, respectively. Taxon and group specific primer sets were then used to detect the presence or absence of native and non-native planktonic life-history stages (propagules) from environmental samples of ballast water and plankton tow net samples collected in Puget Sound. This methodology provides an inexpensive and efficient way to test the discriminatory ability of taxon specific oligonucleotides (PCR primers) before creating molecular probes or beacons for use in molecular ecological applications such as probe hybridizations or microarray analyses. This work addresses the current need to develop molecular tools capable of diagnosing the presence of planktonic life-history stages from non-native marine species (potential invaders) in ballast water and other environmental samples.     

Evaluation of Gambierdiscus survival after exposure to ballast water

The dinoflagellate Gambierdiscus was exposed to ballast water from a trans-oceanic vessel, and maintained at a variety of temperatures in the dark to determine if viability would be maintained. Logarithmically growing Gambierdiscus inocula were admixed (1:6, vol:vol) with ballast water, maintained in the dark at 22.6 °C, 24.6 °C, 26.8 °C and 29.0 °C and assessed for numerical abundance over six days. Calculated growth rates from the biomass time series showed no indication that ballast water negatively impacted Gambierdiscus viability; accompanying microscopic inspections supported this conclusion. Filtration of large volumes of collected ballast water failed to show the presence of any Gambierdiscus cells contained therein. Recovery and microscopic examination of the experimental inocula after 10 weeks in the dark, failed to show cyst development at any temperature regime. This examination of ballast water showed no evidence of cytotoxicity to Gambierdiscus spp.     

Zooplankton distribution and diversity in a frontal area of the Aegean Sea

This study was carried out in the northern Aegean Sea during late summer and spring. The aim was to examine the spatial and temporal distribution of the zooplankton community across a dynamic frontal area and to investigate how the oceanographic heterogeneity structures the composition of the zooplankton assemblages. The low-salinity and cold Black Sea water coming from the Dardanelles Strait is modified by mixing with the underlying warm and saline Aegean water. These hydrological features result in a pronounced thermohaline front in the northern Aegean Sea throughout the year. In both seasons, zooplankton was collected using both 45 and 200 µm mesh plankton nets. A high abundance of zooplankton was observed in the surface layer at the stations closest to the Dardanelles Strait on the stratified side of the front. The zooplankton distribution and community structure in the northern Aegean Sea were strongly influenced by the hydrological features. The frontal structure acts as a boundary for the zooplankton community. The surface layer at the stratified stations had the lowest copepod diversity, from where it increased with depth and horizontally as the stratification weakened outside of the front. The total abundance of zooplankton collected with the 200 µm net was between two and 20 times lower than samples taken with the 45 µm net. The most pronounced differences were observed for the adults and copepodids of the small genera Oithona, Oncaea and Microsetella. Thus, to manage and understand the transfer of primary production up the food chain in the Aegean Sea, the smaller fraction of copepods should be taken into account in future investigations.     

Characterization of bacteria in ballast water using MALDI-TOF mass spectrometry

To evaluate a rapid and cost-effective method for monitoring bacteria in ballast water, several marine bacterial isolates were characterized by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Since International Maritime Organization (IMO) regulations are concerned with the unintended transportation of pathogenic bacteria through ballast water, emphasis was placed on detecting species of Vibrio, enterococci and coliforms. Seawater samples collected from the North Sea were incubated in steel ballast tanks and the presence of potentially harmful species of Pseudomonas was also investigated. At the genus-level, the identification of thirty six isolates using MALDI-TOF MS produced similar results to those obtained by 16S rRNA gene sequencing. No pathogenic species were detected either by 16S rRNA gene analysis or by MALDI-TOF MS except for the opportunistically pathogenic bacterium Pseudomonas aeruginosa. In addition, in house software that calculated the correlation coefficient values (CCV) of the mass spectral raw data and their variation was developed and used to allow the rapid and efficient identification of marine bacteria in ballast water for the first time.     

A comparison of pump sampling systems for live zooplankton collection

Simultaneous and sequential tests of pump sampling equipment showed that sampling mortality of Lake Michigan zooplankton collected from the cooling water intake of a nuclear power plant was dependent on the sampling method used. Two sampling devices and two pump types were tested to determine which combination resulted in the lowest sampling mortality. An in-line filter trap located on the suction side of the pump provided the lowest sampling mortality. Higher sampling rates were achieved with a centrifugal pump without increasing sampling mortality.