Factors affecting preference responses of the freshwater ciliate Uronema nigricans to bacterial prey

ABSTRACT. To enhance our understanding of the factors affecting feeding selectivity of bacterivorous protists in aquatic systems, we examined the preference responses of the freshwater ciliate Uronema nigricans towards three bacterial prey taxa, Pseudomonas luteola, Serratia rubidaea , and Aeromonas hydrophila . Potential factors influencing the predator–prey contact rate included the previous feeding history of the ciliate and physiological state of bacteria. Preference indexes were obtained from multiple-choice mazes in which ciliates moved preferentially towards alternative bacteria or the prey species on which they had been feeding. Uronema nigricans showed differential attraction towards the offered prey types, and these preferences varied as a function of the ciliate feeding history: U. nigricans growing on P. luteola showed lower preference responses towards the offered bacteria than U. nigricans growing on S. rubidaea . The bacteria in stationary phase elicited a higher degree of attraction than bacteria in exponential phase, probably due to a higher concentration of carbohydrates in the former. Therefore, this protist will preferentially swim towards bacteria in stationary growth phase, although the degree of this response will be affected by the recent feeding history of the ciliate.     

Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio

The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations.     

Association of the abundance and vertical distribution of tuna and beakfish in the southeast of the Caribbean sea

The longline hooks suspension depth was estimated using the Mechanic Imitation of Flexible Systems method. The vertical distribution of tunas and billfish was determined by the relative abundance index, obtained from the catch by 11 to 25 m -long longline vessels, -based at Cumaná, Venezuela, South-eastern Caribbean Sea in depths of 65 to 142 m. The CPUE was evaluated per species, according to depth. High values were found for most of the captured species in the layer from 105 to 125 m. Yellowfin tuna (Thunnus albacares) showed the highest yield (3.37 fish/100 hooks) and blue marlin (Makaira nigricans) the lowest (0.04 fish/100 hooks). However, the statistical comparison did not allow to reject the hypothesis of lack of depth efect (Kruskal-Wallis p > .05), and demonstrated a homogeneous distribution of yellowfin tuna (Thunnus albacares), albacore (Thunnus alalunga), bigeye tuna (Thunnus obesus), sailfish (Istiophorus albicans), white marlin (Tetrapturus albidus) and blue marlin (Makaira nigricans) in the water column. The conclusion is that fish concentration in the Southern border of the Caribbean Sea is possibly due to several hydroclimatic factors--which affect tuna and billfish catching--such as water temperature and dissolved oxygen concentration which limit the distribution according to depth.     

Osmoregulatory capabilities of the gray snapper,Lutjanus griseus: salinity challenges and field observations

We investigated the osmoregulatory responses (plasma osmolality and blood hematocrit) displayed by the gray snapper 6–192?h after abrupt changes in ambient salinity. Fish were challenged with six different salinity treatments including a control (0, 5, 30, 50, 60, and 70?ppt) and blood samples were collected at various time points post-transfer. Gray snapper across all size classes tested (13.5–24.5?cm total length) acclimated successfully to hypo- and hyper-saline environments (0–60?ppt) after an adjustment period of ～96?h. However, abrupt transfers to 70?ppt resulted in 100% mortality within 48?h. Laboratory results were then compared with field measurements obtained after fish were captured in low salinity (0–4?ppt) or marine (～30?ppt) habitats, suggesting that osmoregulatory processes occurred similarly in both settings. Overall, findings suggest that gray snapper possess similar or higher osmoregulatory capabilities compared to many euryhaline species examined to date, and thus should be considered a euryhaline species.     

The Cultivation of Symbiote-free Marine Ciliates in Axenic Medium

SYNOPSIS. Media have been developed for axenic cultivation of 10 strains belonging to 7 species of small marine ciliates. A medium containing cerophyl extract, proteose peptone, trypticase, yeast nucleic acid, biotin, calcium pantothenate folic acid, nicotinamide, pyridoxal HCl, riboflavin, thiamine HCl, and DL-thioctic acid in sea water supports the growth of Uronema nigricans strains Pc and 34/2, Parauronema virginiatum strains 2/1 and 19/1, Miamiensis avidus strains Ma and 19/3, Miamiensis sp. strain 1/1, a strain of "G" ciliate, and strains 33/8 and 34/7 of 2 unidentified species. By substituting a mixture of asolecithin, cephalin, and Tween 80 for cerophyl in the medium, luxurious growth of all except the strains of the 2 unidentified species can be obtained. A defined medium consisting of 18 amino acids, 5 purine derivatives, 8 vitamins, asolecithin, cephalin, and Tween 80 in synthetic sea water also has been developed for 6 of the strains: M. avidus Ma and 19/3, Miamiensis sp. 1/1, P. virginiatum 2/1 and 19/1, and U. nigricans Pc. In general the ciliates grow best at pH 7.2 in the dark at 27 C in media containing sea water of density = 1.015. Under these conditions maximum populations are reached in 4–5 days and range from several hundred thousand to 3 or 4 × 10⁶ depending upon the strain. Electronmicroscopic observations for the presence of endosymbiotes gave negative results.     

Salinity tolerance of stable fly (Diptera: Muscidae)

Effects of salinity on the survival, growth, and development of stable fly, Stomoxys calcitrans (L.), were investigated in the laboratory. Larvae failed to develop to pupation when reared in media containing a salinity of 40 parts per thousand (ppt) sodium chloride (NaCl). Maximum salinity supporting larval development equaled the salinity of seawater (34 ppt); the larval LC90 was 24.2 ppt. Deleterious effects of high salinity decreased as larvae matured. Six-day-old larvae reared at a salinity of 34 ppt weighed 79% less than controls, compared with a 36% difference in 9-d-old larvae; by pupation, the difference was only 24%. Salinity did not influence the duration of larval, pupal, or adult stages. Survival of pupae was unimpaired despite a slight increase in number of pupal deformities, and normal adults emerged. Eggs were highly tolerant to saline. They hatched at salinity concentrations lethal to larvae; greater than 50% hatch occurred even when eggs were maintained at 80 ppt NaCl. Sensitivity of larvae to salinities close to that of seawater might be important for control of stable flies inhabiting marine areas.     

Cultural characteristics of the marine ciliated protozoan, Uronema marinum Dujardin

The marine hymenostome ciliate Uronema marinum Dujardin was isolated from sediment from Robin Hood's Bay, Yorkshire. Experiments on the growth of U. marinum were carried out in batch cultures in which the ciliate was maintained on a diet of Vibrio sp. The duration of the lag phase varied directly with the age of the inoculum. Factorially designed experiments were used to investigate the effects of salinity and temperature on the rate of logarithmic growth of the ciliate. A response surface analysis of the data indicated that there was interaction between salinity and temperature. Regression equations predicted a minimum doubling time of 2.5 h at 32‰ and 25 °C. The cell volume of U. marinum varied during growth; it reached a maximum at the end of the lag phase and decreased during the logarithmic phase to become minimal in the stationary phase.     

Tolerance of seahorse Hippocampus kuda (Bleeker) juveniles to various salinities

In line with current conservation efforts, some success in the captive breeding of the seahorse Hippocampus kuda (Teleostei: Syngnathidae) has been achieved. To evaluate the salinity tolerance of these hatchery‐bred juveniles, 9‐week‐old H. kuda were transferred without prior acclimatization from ambient full strength seawater (32–33 ppt) to salinities ranging from freshwater to 85 ppt. Survival, growth, and total body water content were determined after 4 and 18 days of exposure. Juvenile H. kuda are able to survive in dilute seawater (15 ppt) for at least 18 days without any compromise in growth (both wet and dry body weight), survival, and total body water. Fish abruptly transferred to freshwater succumbed within 4–24 h, while survival of 5 ppt‐reared fish decreased to ca. 65% in 18 days. Although 10 ppt‐reared seahorses had growth and survival comparable with the control (30 ppt seawater), total body water was significantly elevated indicating reduced adaptability. The upper limit of H. kuda salinity tolerance was 50 ppt. Fish reared at salinities ≥55 ppt succumbed within 24 h. Like several other marine teleosts, growth and survival of juvenile H. kuda tended to peak in diluted seawater salinities of 15 and 20 ppt. These results indicate the possibility of growing hatchery‐bred H. kuda in brackishwater environments.     

An investigation of the tolerance to ammonia of the marine ciliate Euplotes vannus (Protozoa,Ciliophora)

Hydrobiologia - The toxicity of ammonia and its effects on growth inhibition for the marine ciliate, Euplotes vannus, were measured in a conventional open system. Linear regression analysis showed...     

Trends in tuna carbon isotopes suggest global changes in pelagic phytoplankton communities

Considerable uncertainty remains over how increasing atmospheric CO₂ and anthropogenic climate changes are affecting open‐ocean marine ecosystems from phytoplankton to top predators. Biological time series data are thus urgently needed for the world's oceans. Here, we use the carbon stable isotope composition of tuna to provide a first insight into the existence of global trends in complex ecosystem dynamics and changes in the oceanic carbon cycle. From 2000 to 2015, considerable declines in δ¹³C values of 0.8‰–2.5‰ were observed across three tuna species sampled globally, with more substantial changes in the Pacific Ocean compared to the Atlantic and Indian Oceans. Tuna recorded not only the Suess effect, that is, fossil fuel‐derived and isotopically light carbon being incorporated into marine ecosystems, but also recorded profound changes at the base of marine food webs. We suggest a global shift in phytoplankton community structure, for example, a reduction in ¹³C‐rich phytoplankton such as diatoms, and/or a change in phytoplankton physiology during this period, although this does not rule out other concomitant changes at higher levels in the food webs. Our study establishes tuna δ¹³C values as a candidate essential ocean variable to assess complex ecosystem responses to climate change at regional to global scales and over decadal timescales. Finally, this time series will be invaluable in calibrating and validating global earth system models to project changes in marine biota.     

Role of the 4-phosphopantetheinyl transferase of Trichoderma virens in secondary metabolism and induction of plant defense responses

Trichoderma virens is a ubiquitous soil fungus successfully used in biological control due to its efficient colonization of plant roots. In fungi, 4-phosphopantetheinyl transferases (PPTases) activate enzymes involved in primary and secondary metabolism. Therefore, we cloned the PPTase gene ppt1 from T. virens and generated PPTase-deficient (?ppt1) and overexpressing strains to investigate the role of this enzyme in biocontrol and induction of plant defense responses. The ?ppt1 mutants were auxotrophic for lysine, produced nonpigmented conidia, and were unable to synthesize nonribosomal peptides. Although spore germination was severely compromised under both low and high iron availability, mycelial growth occurred faster than the wild type, and the mutants were able to efficiently colonize plant roots. The ?ppt1 mutants were unable of inhibiting growth of phytopathogenic fungi in vitro. Arabidopsis thaliana seedlings co-cultivated with wild-type T. virens showed increased expression of pPr1a:uidA and pLox2:uidA markers, which correlated with enhanced accumulation of salicylic acid (SA), jasmonic acid, camalexin, and resistance to Botrytis cinerea. Co-cultivation of A. thaliana seedlings with ?ppt1 mutants compromised the SA and camalexin responses, resulting in decreased protection against the pathogen. Our data reveal an important role of T. virens PPT1 in antibiosis and induction of SA and camalexin-dependent plant defense responses.     

An approach to determining the sampling effort for analyzing biofilm-dwelling ciliate colonization using an artificial substratum in coastal waters

A new approach to determining sampling effort for analyzing biofilm-dwelling ciliate colonization was studied in the coastal waters of the Yellow Sea, northern China, from May to June 2010. The optimal sample size for evaluating biofilm-dwelling ciliate colonization increased with shortening exposure time, and can be determined according to the probability of recovering those species with a specified cumulative contribution to communities. More slide-replicates were required at a depth of 3 m than at 1 m to recover equivalent proportions of the ciliate communities. For routine colonization dynamics analyses, 10 slide-replicates (175 cm(2)) were sufficient to achieve a 95% probability of recovering those species with a cumulative contribution of >90% to the ciliate communities at a depth of 1 m. These results suggest that 10 slide-replicates immersed at a depth of 1 m may be an optimal sampling strategy for analyzing the colonization dynamics of biofilm-dwelling ciliate communities in marine habitats.     

Entry of Vibrio harveyi and Vibrio fischeri into the viable but nonculturable state

AIMS: Physiological responses of marine luminous bacteria, Vibrio harveyi (ATCC 14216) and V. fischeri (UM1373) to nutrient-limited normal strength (35 ppt iso-osmolarity) and low (10 ppt hypo-osmolarity) salinity conditions were determined. METHODS AND RESULTS: Plate counts, direct viable counts, actively respiring cell counts, nucleoid-containing cell counts, and total counts were determined. Vibrio harveyi incubated at 22 degrees C in nutrient-limited artificial seawater (ASW) became nonculturable after approximately 62 and 45 d in microcosms of 35 ppt and 10 ppt ASW, respectively. In contrast, V. fischeri became nonculturable at approximately 55 and 31 d in similar microcosms. Recovery of both culturability and luminescence of cells in the viable but nonculturable state was achieved by addition of nutrient broth or nutrient broth supplemented with a carbon source, including luminescence-stimulating compounds. Temperature upshift from 22 degrees C to 30 degrees C or 37 degrees C did not result in recovery from nonculturability. CONCLUSIONS: The study confirms entry of V. harveyi and V. fischeri into the viable but nonculturable state under low-nutrient conditions and demonstrates nutrient-dependent resuscitation from this state. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confirms loss of luminescence of V. harveyi and V. fischeri on entry into the viable but nonculturable state and suggests that enumeration of luminescent cells in water samples may be a rapid method to deduce the nutrient status of a water sample.     

Observations on the ultrastructure, mode of infectivity and host range of xenosomes

Xenosomes are infectious bacterial-like symbionts that reside in the cytoplasm of certain strains of the marine hymenostome ciliate, Parauronema acutum. Ultrastructural studies of xenosomes of one such strain, P. acutum, 110/3, reveal that the particles possess a cell envelope whose outer membrane resembles that of Gramnegative bacteria. Unlike typical Gram-negative bacteria, however, xenosomes possess a much reduced ‘murein’ layer. The DNA is not located in a ‘nucleosome’ or ‘nuclear body’ as it is in most bacteria, but may be found dispersed throughout the internal contents of the particle. Xenosomes are capable of infecting several strains of Parauronema spp. and at least one other ciliate of marine origin, Miamiensis avidus, Ma. They are also capable of inhibiting the growth of certain marine ciliate strains, particularly those of the genus Uronema. Infectivity may be related to the fact that the particles are motile. When released from the host protozoan, they may be seen spinning or darting rapidly, suggesting the presence of a motile apparatus. Phosphotungstic acid staining of the released particles show that they do, indeed, possess bacterial-like flagella. The average number of flagella per particle observed was two, although some particles were seen with as many as five. A flagellum may occur at almost any point on the surface of the particle and is characteristically wider near its point of insertion into the cell envelope than it is near its tip. During normal growth in axenic culture, xenosomes are actively released into the ambient medium. Particles, thus released, rapidly lose their capacity to infect susceptible hosts, and this capacity parallels their ability to remain motile.     

An approach to determining the sampling effort for analyzing biofilm-dwelling ciliate colonization using an artificial substratum in coastal waters

A new approach to determining sampling effort for analyzing biofilm-dwelling ciliate colonization was studied in the coastal waters of the Yellow Sea, northern China, from May to June 2010. The optimal sample size for evaluating biofilm-dwelling ciliate colonization increased with shortening exposure time, and can be determined according to the probability of recovering those species with a specified cumulative contribution to communities. More slide-replicates were required at a depth of 3 m than at 1 m to recover equivalent proportions of the ciliate communities. For routine colonization dynamics analyses, 10 slide-replicates (175 cm²) were sufficient to achieve a 95% probability of recovering those species with a cumulative contribution of >90% to the ciliate communities at a depth of 1 m. These results suggest that 10 slide-replicates immersed at a depth of 1 m may be an optimal sampling strategy for analyzing the colonization dynamics of biofilm-dwelling ciliate communities in marine habitats.     

Sodium chloride inhibits the growth and infective capacity of the amphibian chytrid fungus and increases host survival rates

The amphibian chytrid fungus Batrachochytrium dendrobatidis is a recently emerged pathogen that causes the infectious disease chytridiomycosis and has been implicated as a contributing factor in the global amphibian decline. Since its discovery, research has been focused on developing various methods of mitigating the impact of chytridiomycosis on amphibian hosts but little attention has been given to the role of antifungal agents that could be added to the host's environment. Sodium chloride is a known antifungal agent used routinely in the aquaculture industry and this study investigates its potential for use as a disease management tool in amphibian conservation. The effect of 0-5 ppt NaCl on the growth, motility and survival of the chytrid fungus when grown in culture media and its effect on the growth, infection load and survivorship of infected Peron's tree frogs (Litoria peronii) in captivity, was investigated. The results reveal that these concentrations do not negatively affect the survival of the host or the pathogen. However, concentrations greater than 3 ppt significantly reduced the growth and motility of the chytrid fungus compared to 0 ppt. Concentrations of 1-4 ppt NaCl were also associated with significantly lower host infection loads while infected hosts exposed to 3 and 4 ppt NaCl were found to have significantly higher survival rates. These results support the potential for NaCl to be used as an environmentally distributed antifungal agent for the prevention of chytridiomycosis in susceptible amphibian hosts. However, further research is required to identify any negative effects of salt exposure on both target and non-target organisms prior to implementation.     

Effect of salinity and body weight on ecophysiological performance of the Pacific white shrimp (Litopenaeus vannamei)

Ecophysiological responses of Litopenaeus vannamei were evaluated as functions of environmental salinity and animal size. Growth rate, routine metabolic rate, limiting oxygen concentration, and marginal metabolic scope were determined for L. vannamei acclimated to, and tested at, salinities of 2, 10, and 28 ppt, all at 28 °C. Routine metabolic rate (RMR), estimated as oxygen-consumption rate per unit body weight for fasted, routinely-active shrimp, was independent of salinity but decreased with increasing shrimp weight. Limiting oxygen concentration for routine metabolism (LOCr) decreased with increased shrimp weight for the 10 and 28 ppt treatments, but not for the 2 ppt treatment. Marginal metabolic scope (MMS = RMR/LOCr) also decreased with increasing shrimp weight and was independent of salinity. Growth rate was significantly less at 2 ppt than at either 10 or 28 ppt, which gave similar growth rates.     

Feeding behavior of yellowfin tuna around two insular regions of the western Atlantic Ocean

The yellowfin tuna is a very abundant tropical tuna species in the western equatorial Atlantic Ocean and an important fishery resource for the Brazilian tuna fleet. In this study we performed stable isotope analysis to better understand the spatial trophodynamics and dietary changes in yellowfin tuna around two insular marine protected areas in Brazil. A total of 65 yellowfin tuna specimens measuring between 47 and 138 cm LT (total length) were sampled around the archipelagos of Fernando de Noronha (FNA; n = 34) and Saint Peter and Saint Paul (SPSPA; n = 31) between July 2018 and September 2019. Bayesian mixing models and generalized additive models were used to investigate the contributions of four different prey items (zooplankton, cephalopods, fish larvae, and flying fish) to yellowfin tuna diet in each area and their potential changes in relation to predator growth. The four prey items were found to have different overall contributions between the two studied areas, with zooplankton being the most important prey in FNA, whereas flying fish was the most relevant prey to the species' diet in SPSPA. Significant changes in the species diet by size were also found, with fish smaller than 90 cm (TL) having a more generalist diet and larger animals relying more on consuming larger and more nutritious prey (i.e., flying fish). Our results suggest that these two marine protected areas play an important role in ocean dynamics, providing important and different foraging grounds for the development of this predator species.     

A new strain of Cryptocaryon irritans from the cultured olive flounder Paralichthys olivaceus

An obligate parasite, Cryptocaryon irritans, which is responsible for the white spot disease of marine fish is known to develop in the temperature regime over 19 degrees C. Recently, however, we found white spot disease of olive flounder Paralichthys olivaceus during winter at water temperatures ranging between 12 and 16 degrees C in Korea. In the present study we isolated a C. irritans-like ciliate from the affected fish and investigated its reproductive characters to compare the newly found ciliate with typical C. irritans. The newly found ciliate had an additional process in the reproductive stage, characterized by a budding before palintomic division, and it showed a higher ability to carry out tomitogenesis at a low temperature (16 degrees C) than at a high temperature (24 degrees C). Nevertheless, the present ciliates still had much in common with typical C. irritans with respect to clinical, histopathological, and morphological characters, suggesting that it is a new strain of C. irritans, adapted to lower water temperature.     

Tolerance of soil flagellates to increased NaCl levels

The ability of heterotrophic flagellates to survive and adapt to increasing salinities was investigated in this study. Whole soil samples were subjected to salinities corresponding to marine conditions and clonal cultures were used to perform growth and adaptation experiments at a wide range of different salinities (0-50 ppm). More morphotypes tolerant to elevated NaCl levels were found in road verge soil that was heavily exposed to de-icing salt than in less exposed soils, though there were fewer tolerant than intolerant morphotypes in all soils examined. Heterotrophic flagellates isolated on a freshwater medium from a non-exposed soil were unable to thrive at salinities above 15 ppt, and showed reduced growth rates even at low salt salinities (1-5 ppt). The findings suggest that heterotrophic soil flagellates are less tolerant to NaCl than their aquatic relatives, possibly due to their long evolutionary history in soil, and support the idea that identical morphospecies may differ considerably with respect to physiology