Trade-offs of lipid remodeling in a marine predator-prey interaction in response to phosphorus limitation.

Phosphorus (P) is a key nutrient limiting bacterial growth and primary production in the oceans. Unsurprisingly, marine microbes have evolved sophisticated strategies to adapt to P limitation, one of which involves the remodeling of membrane lipids by replacing phospholipids with non-P-containing surrogate lipids. This strategy is adopted by both cosmopolitan marine phytoplankton and heterotrophic bacteria and serves to reduce the cellular P quota. However, little, if anything, is known of the biological consequences of lipid remodeling. Here, using the marine bacterium Phaeobacter sp. MED193 and the ciliate Uronema marinum as a model, we sought to assess the effect of remodeling on bacteria-protist interactions. We discovered an important trade-off between either escape from ingestion or resistance to digestion. Thus, Phaeobacter grown under P-replete conditions was readily ingested by Uronema, but not easily digested, supporting only limited predator growth. In contrast, following membrane lipid remodeling in response to P depletion, Phaeobacter was less likely to be captured by Uronema, thanks to the reduced expression of mannosylated glycoconjugates. However, once ingested, membrane-remodeled cells were unable to prevent phagosome acidification, became more susceptible to digestion, and, as such, allowed rapid growth of the ciliate predator. This trade-off between adapting to a P-limited environment and susceptibility to protist grazing suggests the more efficient removal of low-P prey that potentially has important implications for the functioning of the marine microbial food web in terms of trophic energy transfer and nutrient export efficiency.

Impacts of inorganic nutrients on the physiology of a mixoplanktonic ciliate and its cryptophyte prey.

Many marine planktonic ciliates retain functional chloroplasts from their photosynthetic prey and use them to incorporate inorganic carbon via photosynthesis. While this strategy provides the ciliates with carbon, little is known about their ability to incorporate major dissolved inorganic nutrients, such as nitrogen and phosphorus. Here, we studied how ciliates respond to different concentrations of dissolved inorganic nitrogen and phosphorus. Specifically, we tested the direct and indirect effects of nutrient availability on the ciliate Strombidium cf. basimorphum fed the cryptophyte prey Teleaulax amphioxeia. We assessed responses in the rates of growth, ingestion, photosynthesis, inorganic nutrient uptake, and excretion. Our results show that the prey changed its carbon content depending on the nutrient concentrations. Low inorganic nutrient concentrations increased S. cf. basimorphum growth and prey ingestion. The higher carbon content of the prey under these low nutrient conditions likely supported the growth of the ciliate, while the higher carbon:nutrient stoichiometry of the prey led to the higher ingestion rates. The low carbon content of the prey at high nutrient concentrations resulted in reduced growth of S. cf. basimorphum, which indicates that carbon acquired via photosynthesis in the ciliate cannot compensate for the ingestion of prey with low carbon content. In conclusion, our findings show S. cf. basimorphum is not able to utilize dissolved inorganic nitrogen and phosphorus for growth, and this species seems to be well adapted to exploit its prey when grown at low nutrient conditions.

Pathological findings and husbandry management in captive Chrysaora spp. medusae affected by umbrellar ulcerative lesions.

During a 12 month period, a group of 14 medusa-stage jellies of the genus Chrysaora, including Pacific sea nettle (Chrysaora fuscescens, n = 11) and Japanese sea nettle (Chrysaora pacifica, n = 3), that were maintained in a public aquarium developed progressive ulcerative umbrellar lesions. In 6 cases (42.9%), ulceration was deep, transmural, and perforated through the mesoglea and subumbrella. In 6 cases (42.9%), ciliated protozoa histomorphologically consistent with scuticociliates were observed in the mesoglea and gastrovascular cavity. In 2 cases (14.3%), commensal dinoflagellates (zooxanthellae) were in the mesoglea and in the cytoplasm of the scuticociliates. During this period, water quality parameters including temperature [°C], pH, oxidation-reduction potential (ORP) [mV], salinity [psu], dissolved oxygen [%], ammonia (NH3), and nitrite(NO2) levels were monitored daily or weekly. The main water quality abnormalities were increased NO2 and pH levels above recommended reference ranges for C. fuscescens and elevated temperature above recommended reference ranges for C. pacifica tank. After correction of water quality parameters, apparent improvement of jellies was observed. In this case, environmental factors were considered the most likely predisposing factors for the development of ulcerative lesions, and ciliated protozoa were considered secondary rather than primary pathogens.

Hysteretic response of Microbial Eukaryotic Communities to Gradually Decreased Nutrient Concentrations in Eutrophic Water.

External environments to microbial eukaryotic communities often change gradually with time. However, whether the responses of microbial eukaryotic communities to these gradually changed environments are continuous or hysteretic and the mechanisms underlying these responses are largely unknown. Here, we used a microcosm to investigate the temporal variation of microbial eukaryotic communities with the gradually decreased nutrient concentrations (nitrogen and phosphorus). We found the differences of microbial eukaryotic community composition and species richness between the control and treatment groups were low during the days 0 to 12, although the nutrient concentrations decreased rapidly during this period in treatment group. However, these differences were clear during the days 14 to 18, although the nutrient concentrations decreased slowly during this period in treatment group. The mechanisms for these results are that the strong homogenous selection (perhaps due to the biotic factors) during the days 8 to 10 in treatment group might enhance the stability of microbial eukaryotic communities. However, the continuously decreased nutrient concentrations weakened the homogenous selection and promoted the strength of environmental filtering, and therefore resulted in the distinct change of microbial eukaryotic communities during the days 14 to 18 in treatment group. Fungi, Chlorophyta and Chrysophyta which associated with the nutrient removal played important roles in this hysteretic change of microbial eukaryotic communities. Overall, our findings suggest that disentangling the non-linear response of communities to gradual environmental changes is essential for understanding ecosystem restoration and degradation in future.

Effects of warming and nutrients on the microbial food web in shallow lake mesocosms.

We analysed changes in the abundance, biomass and cell size of the microbial food web community (bacteria, heterotrophic nanoflagellates, ciliates) at contrasting nutrient concentrations and temperatures during a simulated heat wave. We used 24 mesocosms mimicking shallow lakes in which two nutrient levels (unenriched and enriched by adding nitrogen and phosphorus) and three different temperature scenarios (ambient, IPCC A2 scenario and A2+%50) are simulated (4 replicates of each). Experiments using the mesocosms have been running un-interrupted since 2003. A 1-month heat wave was imitated by an extra 5 °C increase in the previously heated mesocosms (from 1st July to 1st August 2014). Changes in water temperature induced within a few days a strong effect on the microbial food web functioning, demonstrating a quick response of microbial communities to the changes in environment, due to their short generation times. Warming and nutrients showed synergistic effects. Microbial assemblages of heterotrophic nanoflagellates and ciliates responded positively to the heating, the increase being largest in the enriched mesocosms. The results indicate that warming and nutrients in combination can set off complex interactions in the microbial food web functioning.

Environmental dynamics of red Noctiluca scintillans bloom in tropical coastal waters.

An intense bloom of red Noctiluca scintillans (NS) occurred off the Rushikulya estuarine region along the east coast of India, an important site for mass nesting events of the vulnerable Olive Ridley sea turtle. At its peak, densities of NS were 3.3×10(5) cells-l(-1), with low relative abundance of other phytoplankton. The peak bloom coincided with high abundance of gelatinous planktivores which may have facilitated bloom development by their grazing on other zooplankton, particularly copepods. Ammonium concentrations increased by approximately 4-fold in the later stages of bloom, coincident with stable NS abundance and chlorophyll concentrations in the nano- and microplankton. This increase likely was attributable to release of intracellular ammonium accumulated through NS grazing. Dissolved oxygen concentrations decreased in sub-surface waters to near hypoxia. Micro-phytoplankton increasingly dominated chlorophyll-a biomass as the bloom declined, with diminishing picoplankton abundance likely the result of high predation by the ciliate Mesodinium rubrum. Together, these data illustrate factors that can disrupt ecosystem balance in this critically important Indian coastal region.

Phosphorus, nitrogen and chlorophyll-a are significant factors controlling ciliate communities in summer in the northern Beibu Gulf, South China Sea.

Ciliates (protozoa) are ubiquitous components of plankton community and play important roles in aquatic ecosystems in regards of their abundance, biomass, diversity and energy turnover. Based on the stratified samples collected from the northern Beibu Gulf in August 2011, species composition, abundance, biomass, diversity and spatial pattern of planktonic ciliates were studied. Furthermore the main environmental factors controlling ciliate communities were determined. A total of 101 species belonging to 44 genera and 7 orders (i.e., Oligotrichida, Haptorida, Euplotida, Sessilida, Pleurostomatida, Scuticociliatida and Tintinnida) were identified. The variation of ciliate communities was significant at horizontal level, but that was not at vertical level. Based on cluster analysis, ciliate communities were divided into three main groups. Redundancy analysis (RDA) revealed that Group A, existing in the waters with higher concentration of phosphorus and nitrogen, was dominated by Tintinnidium primitivum. Group B in the waters with lower temperature and chlorophyll-a concentration, was dominated by Leegaardiella ovalis. Group C, existing in the waters with higher temperature and chlorophyll-a concentration, was dominated by large Strombidium spp. and Mesodinium rubrum. Combining multiple analytic methods, our results strongly supported that phosphorus, nitrogen and chlorophyll-a were the most significant factors affecting the ciliate communities in the northern Beibu Gulf in summer. Concentration of phosphorus and nitrogen primarily influenced ciliate biomass, implying a potential impact of eutrophication on ciliate growth. The correlation with chlorophyll-a concentration, on one hand indicate the response of ciliates to the food availability, and on the other hand, the ciliates containing chloroplasts or endosymbionts may contribute greatly to the chlorophyll-a.

Impacts of restoration of an uncontrolled phosphogypsum dumpsite on the seasonal distribution of abiotic variables, phytoplankton, copepods, and ciliates in a man-made solar saltern.

The restoration of an uncontrolled phosphogypsum landfill was investigated for its effects on the seasonal distribution of phytoplankton, ciliates, and copepods. Sampling was carried out monthly from September 2007 to August 2008 at four ponds of increasing salinity (A1, 41 psu; A5, 46 psu; A16, 67 psu; and C31, 77 psu) in the Sfax solar saltern (southeastern Tunisia). Physicochemical and biological analyses were carried out using standard methods. Results showed drastic reduction of phosphate input and greater diversity of phytoplankton, ciliates, and copepods than before restoration. Pennate diatoms and new ciliates, considered bio-indicators of less-stressed marine ecosystems, proliferated in the A1 pond for the first time after restoration. Copepods appeared to feed on a wide range of prey. Economically, removal of the 1.7 million m(3) of phosphate improved the quality of the site's salt production, enabling the salt company to receive the quality ISO 9001 accreditation.

Impacts of an uncontrolled phosphogypsum dumpsite on summer distribution of phytoplankton, copepods and ciliates in relation to abiotic variables along the near-shore of the southwestern Mediterranean coast.

In connection with the Taparura Project, studies of spatial distribution of the crustacean zooplankton community, nutrients, phytoplankton and ciliates were conducted in July 2007 at 45 stations spread over fifteen transects along the coast north of Sfax. The results showed that the N/P ratio was lower than the Redfield ratio, suggesting potential N limitation. Phytoplankton was characterised by the proliferation of several diatoms, while ciliates were largely dominated by spirotrichs. Copepods were the most abundant zooplankton present during the entire study period, comprising 61% of the total zooplankton community. Twelve copepod families were identified at every station, with a high percentage of Oithonidae (77% of copepods) dominated by Oithona nana. The abundance of this species was correlated with that of diatoms, Cocoolithophorideae and ciliated Colpodea, suggesting that O. nana may feed on a wide range of prey. Despite human pressure and industrial activities, the coastal waters north of Sfax showed a wide diversity of phytoplankton, ciliates and zooplankton.

Effects of dietary n-3 highly unsaturated fatty acids on growth, nonspecific immunity, expression of some immune related genes and disease resistance of large yellow croaker (Larmichthys crocea) following natural infestation of parasites (Cryptocaryon irritans).

The study was conducted to investigate the effects of dietary n-3 highly unsaturated fatty acid (n-3 HUFA) on growth, nonspecific immunity, expression of some immune related genes and disease resistance of juvenile large yellow croaker (Larmichthys crocea) following natural infestation of parasites (Cryptocaryon irritans). Six isoproteic and isolipidic diets were formulated with graded levels of n-3 HUFA ranging from 0.15% to 2.25% of the dry weight and the DHA/EPA was approximately fixed at 2.0. Each diet was randomly allocated to triplicate groups of fish in floating sea cages (1.0 × 1.0 × 1.5 m), and each cage was stocked with 60 fish (initial average weight 9.79 ± 0.6 g). Fish were fed twice daily (05:00 and 17:00) to apparent satiation for 58 days. Results showed that moderate n-3 HUFA level (0.98%) significantly enhanced growth compared with the control group (0.15% HUFA) (P < 0.05), while higher n-3 HUFA levels (1.37%, 1.79% and 2.25%) had detrimental effects on the growth though no significance was found (P > 0.05). Nitro blue tetrazolium (NBT) positive leucocytes percentage of head kidney and serum superoxide dismutase (SOD) activity increased with increasing n-3 HUFA from 0.15% to 0.60%, and decreased with further increase of n-3 HUFA from 0.60% to 2.25% (P < 0.05). Serum lysozyme activity increased significantly as n-3 HUFA increased from 0.15% to 1.37%, and then decreased with n-3 HUFA from 1.37% to 2.25% (P > 0.05). There were no significant differences in phagocytosis index (PI) of head kidney leucocytes among dietary treatments (P > 0.05). The hepatic mRNA expression of Toll-like receptor 22 (TLR22) and Myeloid differentiation factor 88 (MyD88) was significantly up-regulated in fish fed the diets with low or moderate levels, while in kidney this increment was only found at specific sampling time during the natural infestation of parasites. The 13 d cumulative mortality rate following natural infestation of parasites decreased with n-3 HUFA increased from 0.15% to 0.60% (P < 0.05), and significantly increased with n-3 HUFA from 0.60% to 2.25% (P < 0.05). Results of this study suggested that fish fed low or moderate dietary n-3 HUFA had higher growth, nonspecific immune responses, expression levels of some immune related genes and disease resistance of large yellow croaker following natural infestation of parasites and dietary n-3 HUFA may regulate fish immunity and disease resistance by altering the mRNA expression levels of TLR22 and MyD88.

Diseases of American lobsters (Homarus americanus): a review.

The American lobster fishery is a significant economic driver in coastal communities of North America. Increasingly, the impacts of infectious disease are recognized as important components and factors in the population ecology and subsequent management of the lobster fishery. Both environmental and anthropogenic factors impact marine diseases. The review herein highlights aspects of several important bacterial, fungal and protistan diseases, including gaffkemia, shell disease, vibriosis, disease caused by species of Lagenidium, Haliphthoros and Fusarium, paramoebiasis and Bumper Car disease. As the global environment continues to change, these diseases could more severely affect both wild caught and impounded lobsters.

Relationship of protozoan biomass to phosphate and nitrate removal from activated sludge mixed liquor.

The relationship between protozoan biomass concentration and phosphate and nitrate removal was investigated in mixed liquor using three different carbon sources as supplements. The study was carried out using three respective initial biomass concentrations in a shaking flask environment. Samples were taken every 24 h to determine phosphate, nitrate, dissolved oxygen and chemical oxygen demand. The results revealed a direct relationship between decreases in nutrient concentrations and increases in cell densities of the isolates. Between 24 and 96 h, the increases in the protozoan density corresponded to a phosphate decreases from initial ranges of 55.42-57.36 mg/L, 50.27-51.17 mg/L and 50.01-50.83 mg/L to final ranges of 2.46-11.90 mg/L, 0.61-11.80 mg/L and 1.29-13.89 mg/L, in the presence of Aspidisca, Trachelophyllum and Peranema, respectively. Nitrate concentrations were observed to decrease from initial ranges of 23.84-25.90 mg/L, 23.94-25.84 mg/L and 26.12-26.54 mg/L to final ranges of 0.11-6.32 mg/L, 0.16-5.60 mg/L and 0.24-9.04 mg/L, respectively. The study had revealed that an increase in cell density of the test isolates produces a corresponding increase in phosphate and nitrate removal.

Ciliate communities in a constructed mangrove wetland for wastewater treatment.

In wetlands constructed for treating municipal and industrial wastewater, including mangroves, the effect of wastewater discharged on the substrate has often been neglected. Ciliates, an important group of protozoa, are sensitive to pollutants and any changes in ciliate diversity and community structure reflects the habitat quality. The ciliate communities at six sections along a constructed mangrove belt (33 m in length) planted with Aegicerascorniculatum were investigated in Shenzhen, South China. In all samples collected in both rainy and dry seasons, 183 ciliate species were observed. Most species (56%) were free-swimming forms, while only 10.8% were sessile ciliates. The abundance and species number of ciliates were both found to decrease from the anterior (the wastewater inlet) to the posterior (the outlet) parts of the wetland belt, indicating that organic matter and bacteria in wastewater, which served as food for most ciliates, were gradually removed by the constructed wetland. The r/K (number of r- and K-selected species) ratios at the six sections were relatively small, between 0.2 and 0.4, whereas the C/P (abundance of colpodids and polyhymenophorans) quotient at some sections was higher than 1. These results indicate that although most of the environments along the constructed wetland belt were not stressful for ciliate communities, there were habitats that favored colpodids in high abundances.

Effect of dietary Enterolobium cyclocarpum on microbial protein flow and nutrient digestibility in sheep maintained fauna-free, with total mixed fauna or with Entodinium caudatum monofauna.

Three groups of five wethers with ruminal and duodenal cannulas and maintained as either fauna-free (FF) or inoculated with total mixed fauna (TF) or Entodinium caudatum as a single-species monofauna (EN) were used in an experiment with two 28 d periods. In the first period, the sheep were fed a control barley-based diet (40:60 concentrate to silage DM) and in the second period the diet was supplemented with 187 g DM of Enterolobium cyclocarpum for the last 12 d of the period. The diets of period 1 and 2 were isonitrogenous. There was no effect of fauna on apparent ruminal and total tract organic matter and fibre digestion, but bacterial and microbial N flow and efficiency were improved in FF sheep compared to TF sheep. In period 2, protozoal numbers were reduced between 31 and 88 % 2 h after feeding E. cyclocarpum for the third to twelfth day of supplementation and by an average of 25 % in samples collected over the 24 h feeding cycle. Supplementation of the diet with E. cyclocarpum and the consequent protozoal reduction in TF and EN sheep improved the flow of non-ammonia N and bacterial N to the small intestine and the efficiency of microbial synthesis. However, E. cyclocarpum reduced ruminal organic matter digestion, especially in faunated sheep, and total tract organic matter, N and fibre digestion. Thus, a reduction in the protozoal cell numbers of 25 % was sufficient to achieve the beneficial effects of reduced fauna on the bacterial protein supply, but diet digestibility was reduced.

[An experimental study of potentiated aqueous solutions].

A systematic study was undertaken of luminescent aqueous solutions of homeopathic preparation of sodium chloride at a dilution from D1 to D30, produced by "Weleda" company (Moscow) was carried out. It was shown that intensity of luminescence versus the degree of dilution is a non-monotonous function with several maxima, the main maximum corresponds to 13-14 decimal dilution. The dynamics of spectra was registered for several weeks. A systematic study of water samples (D1-D30) exposed to a similar procedure of potentization but without salt addition was also performed. The difference in the luminescence spectra of water of different stages of potentization was shown. The motility of infusoria Spirostoma ambiquum in solutions being examined was studied. A significant negative correlation between the infusoria motility and luminescence intensity was registered.

Pre- and postweaning attributes in faunated and ciliate-free calves fed calf starter with or without fish meal.

In a 2 x 2 factorial design, 24 newborn, crossbred (Bos indicus x Bos taurus) calves were distributed in 4 equal groups involving dietary treatments of prestarter diets with (FM) or without fish meal (NFM) in a faunated (F) or ciliate-free (D) ruminal environment to study the ruminal fermentative development in pre-and postweaning periods. Defaunation was achieved by rearing calves in isolation and its effect was studied after first appearance of ciliate protozoa (observed after 8 wk of age) in the faunated animals. Calves were fed colostrum for 24 h and whole milk until weaning at 8 wk of age. Ruminal content samples were collected on d 4, 1 wk, weekly to 8 wk, and then biweekly at 9, 11, and 13 wk of age. The samples were analyzed for fermentation products [pH, total volatile fatty acids (VFA) and ammonia N] and enzyme [carboxymethyl (CM) cellulase, xylanase, beta-glucosidase, alpha-amylase, beta-galactosidase, proteases, and urease] activities. Weekly feed intake increased with age, but was similar in both groups. Ruminal pH declined steadily during 0 to 4 wk of age and then stabilized. The total VFA concentration increased with the age. The ammonia N (mg/dL) concentration increased from 14.9 on d 4 to 32.4 at 4 wk, decreased to 17.6 at 8 wk, and then steadied during the postweaning period. Samples collected on d 4 had no fibrolytic activity. Xylanase (U/dL) appeared first (1 wk) followed by beta-glucosidase (U/dL) and CM cellulase (U/dL), which increased steadily from a low of 4.69, 0.08, and 2.95 to 31.8 (6 wk), 5.92 (7 wk), and 19.8 (8 wk), respectively, and the concentrations showed nonsignificant alterations during postweaning periods. The concentration of alpha-amylase (U/dL) increased from 34.3 on d 4 to 87.2 at 8 wk, and then decreased to 56.6 (13 wk). beta-Galactosidase increased up to 6 wk then decreased to trace level (0.20 U/dL) at 13 wk of age. The concentrations of proteases and urease reached a steady state after 1 wk of age. The effect of diet type on ruminal fermentation products and enzyme parameters was nonsignificant. However, a steady and proportional alteration in both parameters in response to dry feed intake with the advancement of age was seen in all calves. Defaunation increased total VFA (97.3 vs. 75.8 mM/L) and alpha-amylase activity (80.3 vs. 61.4 U/dL) and decreased ammonia N (16.4 vs. 21.1 mg/dL), whereas the effect on other parameters was nonsignificant. Ruminal fermentative changes responded to dry feed intake, but did not differ in response to animal protein in prestarter diet.

Coenzyme dependent suppression of 2,2-PDS induced contractures in Spirostomum.

Spirostomum ambiguum was stimulated to contract by addition of the thiol inhibitor, 2,2-dithiobispyridine (PDS), which depletes intracellular reducing equivalents which are generated by coenzyme dependents reactions. PDS induced contractures were not suppressed after 2--4 h of pre-incubation in a medium enriched with ascorbic acid, thiamine, or riboflavin, singly. Two h of incubation with nicotinamide did not suppress contractures; however, 50% suppression occurred after 3 h incubation. Incubation of the cells in a medium enriched with all four coenzymes for up to 4 h, resulted in the suppression of PDS induced contractures to a level as low as 30% of control values. Suppression of contractures by the mixed coenzymes was concentration dependent. Cells that were stimulated with PDS and contracting, exhibited a 50% suppression of contractures within 3 min after transfer to a complementary medium enriched with mixed coenzymes. These results suggest that coenzymes interact synergistically with cellular metabolic processes to inhibit pharmacodynamic responses to PDS.

A kinetic model for microtubule polymerization during oral regeneration in Stentor coeruleus.

The regeneration of oral cilia in the protozoan cell, Stentor coeruleus, has been investigated in the presence of agents which do not favor the polymerization of microtubules. These agents, low temperature and the tubulin binding drugs, podophyllotoxin and beta-peltatin, slow the rate of regeneration. A kinetic model is proposed to test the hypothesis that microtubule polymerization is the rate limiting step in oral regeneration. The results suggest that the tubulin concentration of normal regenerating cells is slightly in excess of an amount which would kinetically limit the regeneration rate.