Hypoxia and seasonal temperature: Short-term effects and long-term implications for Acartia tonsa dana

Far more attention has been given to the short-term lethal impacts of reduced dissolved oxygen on commercially important fish and crabs than to the long-term sublethal impacts on these same species, or on lower trophic levels. This study demonstrates that chronic, sublethal effects of hypoxia on the copepod Acartia tonsa, a critical component of many pelagic coastal food webs, can lead to significant decreases in population growth. The results of laboratory experiments conducted at 15 °C (winter) and 25 °C (summer), under conditions of normoxia (Controls), sublethal hypoxia (1.5 ml l^− 1) and lethal hypoxia (0.7 ml l^− 1) show that egg production female^− 1 day^− 1 was significantly lower at 0.7 ml l^− 1 compared to Controls at both temperatures, while egg production female^− 1 day^− 1 was significantly lower at 1.5 ml l^− 1 compared to controls in both summer experiments and in one of the two winter experiments. Survival was significantly decreased in the 0.7 ml l^− 1 treatment compared to Controls and the 1.5 ml l^− 1 treatment. Copepods developed more slowly and matured at smaller adult body sizes at both temperatures under both lethal and sublethal hypoxia compared to normoxia. Under summer temperatures, egg production was reduced by hypoxia exposure on two counts: (1) exposure to hypoxia during development resulted in smaller adults, which translated into lower egg production, and (2) egg production was still significantly lower in hypoxia treatments compared to Controls even when differences in body size were taken into account. While copepods collected in winter and exposed to winter temperatures and hypoxia also matured at smaller body sizes than copepods exposed to normoxia, egg production in winter was almost entirely attributable to this reduction in body size. These results suggest that coastal hypoxia may have a significantly greater impact in the summer months, when copepod populations are most abundant and growing at their most rapid rate of the year. With the anticipated increases in global temperatures, hypoxia may have even greater impacts on pelagic food webs.     

Effects of estuarine conditions and organic enrichment on the fecundity and hatching success of Acartia clausi in contrasting systems

The fecundity and hatching success of Acartia clausi were analysed at fixed salinity sites (35, 34 and 33 psu) in two nearby estuaries (Bilbao and Urdaibai, Basque coast, Bay of Biscay) from March to June 1997. Field incubations were conducted to estimate egg production rates and hatching success, and the size of eggs and experimental females measured. Water temperature and dissolved oxygen saturation were also determined, as well as seston samples to quantify food abundance and quality. Between-estuary and within-estuary differences were tested statistically, and correlation and regression analyses were used to determine relationships between reproductive and environmental variables. Egg production rates were higher in the organically enriched estuary of Bilbao; this denoting that food supply controls the fertility of A. clausi in these systems. Temporal patterns of egg production differed between estuaries, and were associated with different nutritional factors in each estuary. Within the salinity range analysed, egg production reached higher values at intermediate salinity (≈34 psu) in both estuaries. This was interpreted as the result of the interaction between the positive effect of food increase, and the negative effect of physicochemical conditions with decreasing salinity. Egg size variations mainly occurred temporally in relation to female size, but no clear trade-off between egg size and egg number was observed in any case. A drop in hatching success in Bilbao, mainly in waters of <34 psu, was related to the oxygen depletion caused by organic pollution. This indicates that organic enrichment in Bilbao has opposite effects on the reproductive success, because it enhances egg production but reduces offspring survival.     

Egg production of the copepod Acartia tonsa: The influence of hypoxia and food concentration

Low dissolved oxygen conditions, or hypoxia, occur in estuaries and impact more than just the obvious commercially important species. Copepods are an important link in the food web, and the influence of hypoxia upon them is relatively unstudied. Using the copepod Acartia tonsa, a study of the impact of hypoxia on egg production was conducted. A. tonsa had reduced egg production at low dissolved oxygen concentrations (DO), with the lowest egg production occurring at 0.53-ml/l O₂ concentration. Another experiment was conducted to determine if, by increasing food concentration, the affect of hypoxia could be mitigated. The results indicate that increased food did not offset the impact that hypoxia had on egg production of A. tonsa. These results suggest that as A. tonsa experiences hypoxia in the wild, population numbers will decrease. Thus, if hypoxic conditions increase in scope and duration, declines in copepod abundance may lead to a decline in the abundance of species that depend on them as food. These species may be of commercial importance (e.g., fish, crabs, and oysters).     

Age, distribution and abundance of viable resting eggs of Acartia pacifica (Copepoda: Calanoida) in Xiamen Bay, China

The distribution and abundance of viable resting eggs of copepod Acartia pacifica in Xiamen Bay, China, were determined in the laboratory by the presence of nauplii hatched from the sediments. Sediment cores to a depth of 30 cm, sliced at 1.0 cm intervals, showed that most viable resting eggs of A. pacifica occurred near the sediment surface (0-5 cm), and the number of viable eggs sharply decreased with depth of the sediment, although resting eggs remained viable as deep as 23 cm. ²¹⁰Pb analyses of the sediments indicated that the maximum age of viable eggs of A. pacifica was 20.5 years and the mean egg age was 4.3 years. The egg mortality of A. pacifica in the sediment was 0.1408 year⁻¹, or 85.92% annual egg survival, calculated by regressing ln(egg density) on the age of the sediment. The horizontal distribution of viable resting eggs ranged from 2.27×10³ to 3.85×10⁵ m⁻², with a mean value of 9.49×10⁴ m⁻². Regressions between viable eggs of A. pacifica and all fine-fraction particle size classes (at 2 μm intervals) were not significant. The accumulation of viable resting eggs that can persist for an extended period of time provided evidence for the existence of an egg bank of A. pacifica in the seabed of Xiamen Bay.     

Effects of salinity, temperature and individual variability on the reproduction of Eurytemora affinis (Copepoda; Calanoida) from the Seine estuary: A laboratory study

The brackish water copepod Eurytemora affinis is the most abundant copepod species in the low salinity zone (2-15) of the Seine estuary. Despite its ecological importance, little is known about its population dynamics in the Seine. We studied the effects of temperature (10 °C and 15 °C) and salinity (5, 15 and 25) on reproduction under non-limiting food conditions. We used experiments to determine multiple reproductive parameters for E. affinis. In all experiments, we fed E. affinis a mixture of Rhodomonas marina and Isochrysis galbana. Couples of pre-adult females (C5) and adult males were mated until the female extruded a clutch of eggs and then individual females were observed every 6-12 hours until death to determine (a) embryonic development time, (b) inter clutch time and (c) clutch size throughout their adult lifespan. All reproductive parameters were negatively affected by low temperature (10 °C) and by high salinity (25). At 10 °C and a salinity of 25, mortality during the post-embryonic period was extremely high (85%). Differences in all reproductive parameters between salinities 5 and 15 were minimal. From 15 °C to 10 °C mean latency time (time between hatching of eggs and extrusion of new ones) increased from 0.8 to 2.25 days, the mean embryonic development time from 2.2 to 3.2 days and the mean clutch size decreased from 38 to 22 eggs female^- 1. The mean clutch size decreased when females reached a critical age. The hatching success was high (near 95%) under all conditions except at high salinity. Egg production rates showed no significant differences between salinities 5 and 15 and were significantly higher at 15 °C (13 eggs female^- 1 day^- 1 at salinity 5 and 15) than at 10 °C (4 eggs female^- 1 day^- 1). These values at 15 °C were higher compared to those from other populations of E. affinis in estuaries or lakes. The high reproductive potential of E. affinis from the Seine estuary at 15 °C and low salinities explain its high densities in the low salinity zone during spring and early summer.     

Impact of hypoxia on the survival, egg production and population dynamics of Acartia tonsa Dana

Concern for the increasing occurrence of coastal zone hypoxia has generally focused on the direct, short-term impact of reduced dissolved oxygen (DO) concentrations on the survival of commercially important species such as fish and crabs. Copepods, especially the naupliar stages, are important pelagic food web components, yet only a few studies have considered the effect of reduced DO concentrations on their survival and population dynamics. This study considers the impact of both lethal and sublethal DO concentrations on copepods. Acartia tonsa were reared at 25 °C at saturating DO (normoxic control) and reduced (hypoxic) DO concentrations of 1.5 or 0.7 ml l⁻¹. Oxygen concentrations were maintained in replicate flasks, by bubbling seawater with air (control), or mixtures of nitrogen and oxygen. Egg production, but not survival, was significantly higher in the controls compared to the 1.5 ml l⁻¹ DO treatment. Survival and egg production were significantly lower at 0.7 ml l⁻¹ DO compared to the control. The results suggest that the sublethal as well as the lethal effects of hypoxia may have important repercussions on population and community dynamics in coastal systems.     

Benthic resting eggs of calanoid copepods in Norwegian enclosures used in mariculture: abundance, species composition and hatching

Abundance, species composition and viability of benthic resting eggs of calanoid copepods were investigated in the sediments of seven Norwegian enclosed pond systems used in mariculture as well as in the sediments of the nearby open sea. The ponds represented different types of morphometry and zooplankton communities and were located from approximately 60 to 70 °N. Differences in environmental conditions in deep water also typical with anoxia and the presence of hydrogen sulphide at most of the localities. The lowest and highest egg densities in the enclosures were 7.5 ± 6.4 × 10⁴ and 5.5 ± 2.5 × 10⁶ m^-2, respectively, higher than in the sediments of the nearby open sea. Highest densities were found at larger or median depths. Altogether, egg densities higher than 10³ m^-2 were found of only four calanoid species; Eurytemora affinis, Acartia teclae, Acartia clausi, Temora longicornis. Hatching success was variable (0–92%), and no significant correlation to the presence of anoxia and hydrogen sulphide was found. This high hatching success was achieved with eggs exposed to hydrogen sulphide for several months. The recruitment potential to the plankton of nauplii was estimated from 4.8 to 1682 nauplii l^-1 for the different localities. The results showed that resting egg production is a common life history characteristic of some neritic calanoid species in Norwegian waters. In addition to copepod eggs, low densities (< 10³ m^-2) of eggs from rotifers and cladocerans were found in most of the localities.     

Cost of oviposition site selection in a water strider Aquarius paludum insularis: Egg mortality increases with oviposition depth

Females generally avoid selecting sites for oviposition which have a high predation risk to increase offspring survival. Previous studies have focused on costs to ovipositing females. However, although offspring may also incur costs by being oviposited at low predation risk sites, no studies have focused on costs to offspring. Such costs to offspring were examined by using Aquarius paludum insularis, females of which avoid eggs parasitism by ovipositing at deep sites. Deep sites are safe from egg parasitism but may be unsuitable for hatching due to environmental factors. We examined the costs to offspring at deep sites by comparing the hatching rate, the duration to hatching and the proportion of drowned larvae between eggs that were set at three levels of water depth (0 cm, 25 cm and 50 cm depth). While the hatching rate at 50 cm was lower than that at 0 cm, the rate at 25 cm did not differ from that at 0 cm. Duration to hatching and the proportion of drowned larvae did not differ between the three depths. It is suggested that the declining survival rate of A. paludum eggs was due to increased water pressure at greater depth. Such a cost may exist in other species and such an observation may aid in understanding oviposition site selection.     

Nucleic acid indices of egg production in the tropical copepod Acartia sinjiensis

Two experiments were conducted to evaluate the effects of food and temperature on the nucleic acid content and egg production (and their relationship) of the tropical copepod Acartia sinjiensis. Experiment 1 evaluated the effect of food quality (as different algae species) on the relationship between nucleic acid content and egg production. In Experiment 2, the main and interaction effects of food type, food concentration and temperature on the total, Carbon and Nitrogen specific egg production and nucleic acid indices were evaluated in a factorial experimental design. Food quality, concentration and temperature significantly affected the nucleic acid content, egg production rate and the nucleic acid-egg production relationship of A. sinjiensis. RNA indices were correlated with egg production in females fed Pavlova salina, Tetraselmis chuii and Chaetoceros muelleri, but not in females fed Isochrysis aff. galbana. The slopes of the linear regressions of RNA indices as predictors of egg production were similar in females fed different algae species, suggesting that the slope of these relationships might be independent of food quality. The DNA content of females was significantly affected by food and temperature, suggesting that it is not a good index of cell number in this species. Nevertheless, the RNA:DNA ratio was as good a predictor of egg production as total RNA content. Egg production showed a weakly positive correlation with temperature. On the other hand, total, C- and N-specific nucleic acid indices had a strong negative correlation with temperature. In addition, temperature had a non linear effect on the slopes of the regression lines of RNA content and RNA:DNA ratio as predictors of egg production—slopes were similar at 25 °C and 30 °C, but significantly lower at 20 °C. Furthermore, the predictability of egg production was improved when the interaction term of nucleic acid indices with temperature was used instead of the nucleic acid indices alone in linear regression models. Our results suggest food quality has a limited influence on the nucleic acid-egg production relationship, and that temperature should be accounted for in models using nucleic acid indices as predictors of egg production in A. sinjiensis.     

Effect of essential dietary fatty acids on egg production and hatching success of the marine copepod Temora longicornis

The calanoid copepod Temora longicornis and its food (seston of size < 200 μm) was sampled during three successive seasons (2002-04) in the Trondheimsfjord, Central Norway. Egg production (24 h) and hatching success (72 h) was determined by incubation experiments, and the essential fatty acid (EFA) content of their in situ food was analysed. The dominant EFA in the seston were DHA (22:6n-3) and EPA (20:5n-3), whereas ARA (20:4n-6) were present in low quantities. Egg production and hatching success was relatively low during early spring and late autumn (~ 10 eggs female^- 1 day^- 1 and ~ 30%), and relatively high during summer. Spring phytoplankton, dominated by diatoms, contained low amounts of DHA. Dinoflagellates, small flagellates, and ciliates dominated during summer, when a high content of DHA was recorded.The rate of egg production of T. longicornis did not show any relationship with food concentration (r² = 0.003), but was positively correlated to temperature, although not statistically significantly (r² = 0.48, p = 0.05). The quantitative and percent DHA contents of the food was significantly related to the rate of egg production (r² = 0.96 and 0.95, respectively, p < 0.001), but no such relationship were observed for the quantitative or percent content of EPA and ARA in the seston. The egg production of T. longicornis during May-August was 43-47 eggs female^- 1 day^- 1, with dietary DHA contents higher than 7-8 mg DHA g^- 1 DW. Also the hatching success of T. longicornis was positively correlated to the quantitative content of DHA in the diet (r² = 0.88), but hatching was also inversely related to the percentage ARA (r² = 0.84). The maximum hatching success was found when the ARA content was < 0.15% of total fatty acids and the DHA:ARA ratio was > 50. The conclusion that DHA most strongly affected egg production whereas ARA affected hatching, fit well with earlier findings for fish. Our results do not exclude that toxic aldehydes interact with reproduction of calanoid copepods when diatoms are present, but we observed a consistent pattern where dietary DHA and ARA alone explained a majority of the variability in egg production and hatching of T. longicornis.     

Effects of different diets on the reproduction and naupliar development of the copepod Acartia bifilosa

The influence of diatoms on the reproduction and naupliar development of Acartia bifilosa was investigated under laboratory conditions, comparing initial in situ values and laboratory-food treatments. Egg production by A. bifilosa was significantly reduced by one diatom diet (Phaeodactylum tricornutum: Pt) and by two non-diatom diets (Platymonas subordiformis: Ps and Nannochloropsis oculata: No). It was less affected by the other diatom diet (Skeletonema costatum: Sc) or by two mixed-food treatments (D-mix and DG-mix), composed of two diatoms (Pt, Sc) and four species (Pt, Sc, Ps and No), respectively. The negative effect of Pt was eliminated when adult copepods were offered mixed-food diets. There were no significant differences between the hatching success values observed in filtered seawater and in algal exudates, indicating that diatoms did not produce active dissolved toxic substances under the different food concentrations tested. The mortality rate of nauplii was higher with Pt than the other diets, suggesting that this diatom species had a negative effect on egg production, hatching success and naupliar survival simultaneously. Compared to other diets, No and Pt were not beneficial food sources for reproduction and for female and larval survival.     

Cytotoxicity from sulfide exposure in a sulfide-tolerant marine invertebrate

It is unknown whether sulfide-tolerant marine invertebrates suffer cytotoxicity from sulfide exposure in vivo at environmentally-relevant concentrations. We tested this with the mudflat polychaete Glycera dibranchiata. Exposure of live animals to sulfide up to 2.4 mmol l^− 1 for 24 h did not affect animal survival, and animal condition (gross appearance and activity) was unaffected at sulfide concentrations up to 0.25 mmol l^− 1. However, animal condition was decreased at higher sulfide concentrations (P < 0.0001, n = 33). Coelomic fluid obtained from the animals showed decreased erythrocyte count (P = 0.0035, n = 14), indicating cell loss, and increased propidium iodide and Hoechst 33324 staining (P < 0.0001 and P = 0.0003, respectively, n = 14), indicating loss of plasma membrane integrity, even at sulfide concentrations that produced no change in animal condition. When G. dibranchiata were allowed a 72 h recovery period following 24 h sulfide exposure, there was no overall improvement in condition (P ≥ 0.12, n = 7-8), and worms that had been exposed to 1 mmol l^−  1 sulfide still had an erythrocyte count that was less than half that of control animals (P = 0.028, n = 7). The inability to completely recover the cell count was at least partially due to impaired production of new erythrocytes, since analysis of BrdU incorporation indicated that erythrocyte proliferation was reduced from 2% per day in control animals to 0.12% per day in animals exposed to 1 mmol l^− 1 sulfide (P = 0.010, n = 21). Together, these findings indicate that at least some sulfide-tolerant marine invertebrates experience significant cellular injury and impaired tissue proliferation when exposed to environmentally relevant sulfide concentrations, even when the appearance and behavior of the animal appear unaffected.     

Chitobiase of planktonic crustaceans from South Atlantic coast (Southern Brazil): Characterization and influence of abiotic parameters on enzyme activity

Chitobiase is one of the enzymes involved in chitin degradation in nature. It is produced and released by a variety of organisms from bacteria to fish. In crustaceans, it is associated with digestive function and acts on the epidermis during the molting process. In the present study, the influence of water pH, temperature and salinity on maximum chitobiase activity (MCA), as well as the enzyme affinity (Km) for a substrate, the methylumbelliferyl N-acetyl-ß-d-glucosaminide (MUFNAG) was evaluated in the copepod Acartia tonsa. Km values for chitobiases of other crustaceans from the Patos Lagoon estuary and Cassino Beach (Southern Brazil) were also determined. For A. tonsa, MCA was observed at pH 5-6 and 30-35 °C. The range of pH was quite similar to that reported for other aquatic organisms. However, the range of temperature was lower than that previously reported. For salinity, no previous studies have considered the influence of this parameter on MCA. For A. tonsa, MCA was observed in freshwater, showing a significant linear decrease with increasing salinity. Considering that maximum copepod survival and growth rates are observed between 15 and 25 ppt, these findings suggest that the observed enzyme activity in this range of salinity (68 to 47% of that measured in freshwater) is not a limiting factor for A. tonsa growth. However, the extremely decreased enzyme activity observed in salinity 30 ppt (33% of that measured in freshwater) suggests that chitobiase activity might be one of the limiting factor for copepod growth at 30 ppt salinity or higher. Km values (μM) determined for organisms evaluated in the present study (copepod A. tonsa = 20.77; mysid Metamysidopsis elongata atlantica = 14.67; nauplii barnacle Balanus improvisus = 18.19; decapod zoea = 14.30; decapod megalopa = 24.77) were lower than those reported for other crustaceans from Northern Hemisphere. Also, they were much lower than those of organisms from different taxonomic groups like bacteria and fungi, but much higher than in protozoans and dinoflagelates. These findings suggest that chitobiase might be differentially evolved in each specific group of organism, and even within different ontogenetic stages of the same species, for a better adaptation to cope with its respective environmental needs.     

Potential of an indigenous strain of the entomopathogenic fungus Beauveria bassiana as a biological control agent against the Red Palm Weevil, Rhynchophorus ferrugineus

The potential of a strain of Beauveria bassiana (Ascomycota: Clavicipitaceae) obtained from a naturally infected Rhynchophorus ferrugineus (Coleoptera: Curculionidae) pupa as a biological control agent against this weevil was evaluated both in the laboratory and in semi-field assays. Laboratory results indicate that this strain of B. bassiana can infect eggs, larvae and adults of R. ferrugineus (LC₅₀ from 6.3 × 10⁷ to 3.0 × 10⁹ conidia per ml). However, mortality was not the only indicator of treatment efficacy because adults of either sex inoculated with the fungus efficiently transmitted the disease to untreated adults of the opposite sex, with male-to-female and female-to-male rates of transmission of 55% and 60%, respectively. In addition, treatment with B. bassiana significantly reduced fecundity (up to 62.6%) and egg hatching (32.8%) in pairing combinations with fungus-challenged males, females or both sexes. Likewise, 30-35% increase in larval mortality was observed in larvae obtained from eggs from fungus-challenged females or from untreated females coupled with inoculated males, resulting in an overall 78% progeny reduction. Semi-field preventive assays on potted 5-year old Phoenix canariensis palms, with efficacies up to 85.7%, confirmed the potential of this strain as a biological control agent against R. ferrugineus.     

Nematicidal activity of Haplophyllum tuberculatum and Plectranthus cylindraceus oils against Meloidogyne javanica

The potentials of Haplophyllum tuberculatum and Plectranthus cylindraceus oils to control Meloidogyne javanica were investigated in vitro and in a greenhouse. A mixture of Haplophyllum and Plectranthus oils (1:1) was highly toxic to M. javanica in vitro, as it killed all nematode juveniles and inhibited hatching of eggs at 12.5 μg/ml concentration after 24 h exposure time, as did carbofuran at the same concentration. In the green-house, tomatoes grown in soil treated with a combination (1:1) of the two oils developed fewer root galls than those grown in soil treated with higher doses of either oil. The oil mixture, at 2.5 and 5.0 μg/ml of soil, was not phytotoxic to tomato plants as evident from the appearance and height of plants after 12 weeks exposure time, compared to treatment over the same period at lower effective doses. The nematicidal activity of the combined essential oils was suggested by the presence of C₁₀ dienes, C₁₀ trienes and C₁₀ phenol.     

Toxin content and toxic effects of the dinoflagellate Gyrodinium corsicum (Paulmier) on the ingestion and survival rates of the copepods Acartia grani and Euterpina acutifrons

Short-term experiments showed that ingestion rates of the copepods Acartia grani and Euterpina acutifrons on different concentrations of the dinoflagellate Gyrodinium corsicum were not usually different from those obtained with the non-toxic and similar sized Prorocentrum triestinum. Long-term experiments showed that no A. grani survived for more than 288 h on concentrated cultures (1500 μg C l⁻¹) of G. corsicum compared with survival rates of 86.7% on the non-toxic P. triestinum. In contrast high and similar survival rates were found for E. acutifrons exposed for the same time to similar concentrations of both dinoflagellates. These results demonstrated that G. corsicum produce toxins which have significant adverse effects on the long-term survival rates of some copepods like A. grani but not on other copepods like E. acutifrons. The possibility of PSP-toxin production by G. corsicum was analysed by chromatographic analysis (HPLC-FD) and mouse bioassay of extracts obtained from cultures maintained in exponential growth phase in f/2-Si and L1 culture mediums. These analyses and the bioassay ruled out the presence of PSP toxins in this dinoflagellate. However, mouse tests provided the first evidence for the presence of some type of NSP toxin in G. corsicum. Haemolytic assays also gave the first positive results for the methanol extract of this dinoflagellate species.     

Simultaneous study of particle reworking, irrigation transport and reaction rates in sediment bioturbated by the polychaetes Heteromastus and Marenzelleria

The present study employed simultaneously methods to investigate particle and solute transport and reaction rates in sandy sediments inhabited by two worms (2500 m^− 2) with different feeding modes. Heteromastus filiformis is a head-down deposit-feeder and the main activities exerted by this worm are transport of particles as faecal pellets from subsurface to surface sediments and burrow ventilation. Marenzelleria viridis is a surface deposit-feeder that actively searches for food by burrowing near the sediment surface, producing a network of ventilated galleries in this zone. M. viridis exhibited 1.5 to 2.2 times higher particle mixing rates (D_b = 3.3 to 4 × 10^− 3 cm^− 2 d^− 1) compared to H. filiformis. In M. viridis treatments, continuous advection (eddy diffusion) was the major factor influencing solute transport resulting in apparent diffusion rates (D_a = 2.2 cm^− 2 d^− 1), which were 3 times higher than molecular diffusion within the sediment. In H. filiformis inhabited sediments, the transport of solutes was discontinuous and driven by a surprisingly high nonlocal exchange (α = 1.1-1.3 d^− 1), emphasizing its strong irrigation effects. Accordingly, the enhancement of solute fluxes was more pronounced for H. filiformis compared to M. viridis. Depth integrated TCO₂ production derived from diagenetic modelling, which takes into account three reaction zones, is in good agreement with rates obtained from measured fluxes, indicating the applicability of both approaches to get reliable rates. However, the reaction rates showed that the presence of animals had a modest effect on microbial carbon oxidation. The results proved that transport conditions are deeply related to feeding modes. Exchange of solutes was the most important transport process by H. filiformis, while M. viridis affected both mixing and solute transport.     

X-ray microanalysis (EDXMA) of cadmium-exposed eggs of Bothriocephalus acheilognathi (Cestoda: Bothriocephalidea) and the influence of this heavy metal on coracidial hatching and activity

Over recent years it has been established that pollutants can have a significant impact on host-parasite systems in the aquatic environment, so much so that it has been proposed that parasite fauna may be a useful parameter to monitor water quality. Surprisingly, with perhaps the exception of trematodes and bioaccumulation in adult acanthocephalans, detailed observations on the interaction between helminths, particularly cestodes, and pollutants such as heavy metals, are lacking. In this study, eggs of the carp tapeworm, Bothriocephalus acheilognathi were exposed to a range of cadmium concentrations (0.1, 10, 100 and 10,000 μg/L) and coracidial hatching and survival assessed. Results indicated that the egg is highly resistant to heavy metal pollution and hatching occurs even at 10,000 μg/L. In contrast, the activity of the liberated coracidium significantly decreased after 1 h exposure to cadmium at 10 and 100 μg/L. Electron microscopic X-ray microanalysis of parasite eggs exposed to 1000 and 10,000 μg/L cadmium revealed that cadmium accumulates on the surface of the egg and does not penetrate detectably into the enclosed coracidium. This means that the parasite eggs may be able to withstand a heavy metal pollutant incident.     

Description of Brenneria roseae sp. nov. and two subspecies, Brenneria roseae subspecies roseae ssp. nov and Brenneria roseae subspecies americana ssp. nov. isolated from symptomatic oak

Gram-negative, facultatively anaerobic bacteria were isolated from symptomatic oak tissue in the UK and USA. Partial gyrB sequencing placed ten strains in the genus Brenneria, with B. goodwinii as the closest phylogenetic relative. The strains were investigated further using a polyphasic approach including MLSA (based on partial gyrB, rpoB, infB and atpD gene sequences), 16S rRNA gene sequencing, DNA–DNA relatedness studies and both phenotypic and chemotaxonomic assays. The MLSA and 16S rRNA gene analyses separated the strains into two groups based on origin, suggesting that they belong to Brenneria as two novel species. However, the DNA–DNA relatedness values revealed a closer relationship between the groups and indicated that they should belong to the same species. As the two groups of strains from the UK and USA can be differentiated from each other phenotypically and by ERIC PCR fingerprints, it is proposed to classify them as novel subspecies of a novel Brenneria species. The name Brenneria roseae sp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) is proposed, with Brenneria roseae subsp. roseae ssp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) for the strains from the UK and Brenneria roseae subsp. americana ssp. nov. (FRB 223^T = LMG 27715^T = NCPPB 4582^T) for the strains from the USA.     

Observations on the existence of lower threshold and upper critical food concentrations for the copepod Acartia tons a Dana

Acartia tonsa Dana represents a genus of temperate-tropical inshore copepods which, by virtue of its high biomass and rapid generation times, may be assumed to be an important planktonic primary consumer in terms of total production world-wide; its grazing response to different food concentrations is little known. Using wide ranges of concentrations of phytoplankton cultures, we have found that A. tonsa has a maximum grazing rate of ≈ 10.0 μg chl $\text{a}\text{1}$, decreasing to zero below 1.0 μg chl $\text{a}\text{1}$. This was confirmed using the more limited range of naturally-occurring particulate material. Although grazing rate became progressively reduced above 10 μg chl $\text{a}\text{1}$, ingestion rates continued to increase over the next order of magnitude of food concentration. It appears that A. tonsa is rarely exposed to food concentrations in its natural environment (as measured by conventional techniques) high enough to stimulate the maximum grazing effort. On the other hand, it is suggested that the continued increase of ingestion rates in the laboratory at much higher concentrations may indicate an adaptive mechanism associated with the encountering of ephemeral micro-patches of such concentrations, which would permit rapid filling of an empty gut in an energy-efficient manner. Fecal pellet production was also measured and shown to be a good indicator of ingestion rate.