The effect of temperature on reproduction in three species of cyclopoid copepods

Megacyclops viridis (Jurine), Macrocyclops albidus (Jurine), and Acanthocyclops vernalis (Fischer) were raised in the laboratory at six temperatures (5, 8, 10, 12, 15 and 20°) and fed a mixture of ciliates (Paramecium caudatum and Colpidium campylum). Data were taken on clutch size, embryonic development time, interclutch period, time to first clutch, sex ratio and longevity.Clutch size, time to first clutch, embryonic development time, interclutch period and longevity are inversely related to temperature. The ratio of males to females in a cohort is independent of temperature.     

Regulation of species richness by advection and richness-dependent processes in a coastal fish community

The WWF Oasis of Palo, 38 kms north of Rome, is one of the last remnants of the Mediterranean plain forest formerly covering the coastal areas of Latium. Within this preserved area, nine temporary waterbodies were periodically visited during 2001 to obtain information on their morphology, water cycle and faunal composition. In total, 25 crustacean species were listed and monthly changes in their abundance were recorded. Cladoceran species prevailed in all waterbodies (11 species, 7 genera, 2 families). Copepoda were represented by 8 species, 7 genera, 2 families. Ostracoda included 5 species, 5 genera, 3 families. Anostracans were recorded in only two pools and were represented by Chirocephalus diaphanus Prévost, 1803, the most common species in Italy. Chirocephalus kerkyrensis Pesta, 1936 usually occurring in the waterbodies of the Mediterranean maquis, was missing. Species richness was maximal in late autumn and winter, and correlated positively with pond duration, flooded area, water level and number of rainpools. The species most frequently recorded were Chydorus sphaericus O.F. Müller, 1785 and Daphnia curvirostris Eylmann, 1887 for Cladocera, Megacyclops viridis Jurine, 1820, Diacyclops bicuspidatus odessanus Shmankevitch, 1875 and Mixodiaptomus kupelwieseri Brehm, 1907 for Copepoda, Cyclocypris ovum Jurine, 1820 and Eucypris virens Jurine, 1820 for Ostracoda. Others, like Alona affinis Leydig, 1860, Ceriodaphnia pulchella Sars, 1862 (Cladocera), Macrocyclops albidus Jurine, 1820 (Copepoda), Herpetocypris reptans Baird, 1835 Fabaeformiscandona fabaeformis Fischer, 1854 (Ostracoda) were found in only one pool.     

Ontogenetic Shifts in the Ability of the Cladoceran,Moina macrocopa Straus and Ceriodaphnia cornuta Sars to Utilize Ciliated Protists as Food Source

The ontogenetic diet shifts and age specific ability of the two cladoceran species Moina macrocopa and Ceriodaphnia cornuta to derive energy from ciliated protists have been investigated in laboratory. The postembryonic developmental rates and life table demography (longevity, age and size at first reproduction, fecundity and intrinsic rate of natural increase) of the cladocerans have been elucidated on algae (Chlorella vulgaris) and the ciliated protists (Tetrahymena pyriformis, Colpoda (c.f.) steini) as food. For either of the cladoceran, the somatic growth rate and average body size at first reproduction were higher with algal diet. During initial stages of development (0–5 days), either cladoceran realized higher rate of somatic growth on algal diet, subsequently ciliated protists supported significantly higher growth rate than the alga. Algal and ciliate diets did not differ in maximum body size (C. cornuta: 539–554 μm; M. macrocopa: 1274.8–1309 μm) reached by either of the cladocerans. The maximum body sizes were larger than size at first reproduction with either of the ciliated protists, however, with algal diet the maximum body sizes did not differ from the size at first reproduction in each case. In case of C. cornuta the generation time (20.5 ± 0.3 days on ciliate; 15.6 ± 0.17 days on algal diet), reproductive rates (net reproductive rate: 20.05 ± 3.2 on ciliate; 15.5 ± 1.2 on algal diet), and average life expectancy at hatching (27 ± 0.8 days on ciliate; 22.7 ± 0.71 days on alga) were higher, whereas the size at first reproduction (482 μm on ciliate; 521 μm on alga) was smaller with the ciliate than with an algal diet. The algal and the ciliate diets did not differ in survival (life expectancy at hatching: 9.2 ± 0.7 days) and fecundity (NRR: 23.6 ± 2.4) for M. macrocopa. The two ciliates used in the experiment did not differ in their performance as food source for either cladoceran species. Our results suggest that both the cladoceran species are able to utilize smaller ciliate (e.g., T. pyriformis, C. (c.f.) steini) as food; however with differential ability to derive energy from the ciliate diet and this ability is size and age structured in both cases. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Experimental Approach to the Role of Protozoa in Aquatic Ecosystems

In enrichment batch experiments, samples from three water bodieswere alternatively supplemented by various amounts of organicmaterial and incubated at 20 C. Colpidium campylum reached itshighest total cell volumes in cultures with the highest initialtotal cell volumes in cultures with the highest initial concentrationsof organics; Cyclidium glaucoma preferred lower concentrations;and Glaucoma chattoni occupied the intermediate position. Noneof the species preferred any special type of organic material.In two-stage continuous-flow units, a mixed culture of bacteriawas kept in stage I and the clones of ciliates were maintainedin stage II. The interrelations between the total cell volumesof ciliates at various concentrations of bactopeptone were inaccordance with the results from the enrichment experiments.Since the growth of bacteria continued in the presence of ciliates,a four-stage apparatus was constructed in which a bacterialculture was raised and diluted in three stages before enteringthe culture ofColpidium. The bacterial growth in the presenceof Colpidium was not eliminated even by this arrangement, asdemonstrated by dosing antibiotics along with bacteria. An effectof ciliate metabolites on bacterial growth rate is suggested,completing a metabolic cycle in the bacteria-protozoa system.     

The behavioural responses and tolerance of freshwater benthic cyclopoid copepods to hypoxia and anoxia

The response of four benthic cyclopoid copepods,Acanthocyclops viridis (Megacyclops viridis) (Jurine, 1820),Macrocyclops albidus (Jurine),Eucyclops agilis (Koch, Sars) (Eucyclops serrulatus) (Fischer, 1851) andParacyclops fimbriatus (Fischer), to hypoxia and anoxia was investigated. All of these species died within six hours when confronted by totally anoxic conditions, but all survived four days at oxygen saturation levels as low as 25%. Males succumbed to the effects of anoxia more rapidly than the larger females of each species, and larger species survived for shorter periods than smaller species. In artificially stratified columns, where the lower layer was anoxic, all four species displayed an upward migratory response towards oxygenated conditions. Where the artificial hypolimnion was hypoxic, however, the migratory response was not observed. The results suggest that some benthic copepods cope with seasonal anoxia in eutrophic stratified lakes by migration rather than the various physiological adaptations shown by planktonic and semi-planktonic species.     

Culture of Stentor coeruleus on Colpidium campylum*

SYNOPSIS. A method is described for culturing Stentor coeruleus in agar-lined petri dishes using the smaller ciliate Colpidium campylum as food organism.     

Respiratory studies on two benthic copepods Acanthocyclops viridis and Eucyclops agilis at environmental temperatures

Summary The rate of oxygen consumption in all developmental stages of two benthic cyclopoid copepod species, Acanthocyclops viridis (Jurine) and Eucyclops agilis (Koch, Sars) was investigated at temperatures between 5° C–20° C, which are reflective of the environmental temperature range of the benthic zone in Esthwaite Water, Cumbria, from which the organisms were derived. The larger species, A. viridis was found to have a temperature insensitive metabolism, whereas E. agilis showed a marked temperature response in respiratory function. Gravid and non-gravid females had similar rates of respiration except at 15° C–20° C in E. agilis. Males exhibited much lower respiration rates than females, but higher weight specific respiration rates; these differences were largely attributable to size differentials between the sexes. Estimates of swimming activity were made and these were related to the patterns of respiratory function observed.     

Colpidium-Produced RNA as a Growth Stimulant for Tetrahymena*

SYNOPSIS. During logarithmic growth, Colpidium campylum produced a complex, heat-stable material which elevated the reproductive ate of Tetrahymena subsequently inoculated into the culture. This material stimulated only early stages in the growth of Tetrahymena—effect accompanied by an increased number and size of food vacuoles. The Colpidium product was found to be RNA, precipitable from the medium by acetone. Lipids and proteins or peptides were also present in the complex; these appeared to protect the RNA from the action of chemical and physical agents.     

Taxonomy and ecology of some ciliates (Protozoa,Ciliophora) of the saprobic system. III. Revision of the genera Colpidium and Dexiostoma,and establishment of a new genus,Paracolpidium nov. gen.

The genera Colpidium Stein, 1860 and Dexiostoma Jankowski, 1967 are revised. Our monographic treatment is based on a morphologic and biometric reinvestigation of the main species of these genera and on an evaluation of data from the literature. Colpidium comprises four species, namely C. colpoda (Losana, 1829), C. singulare Vuxanovici, 1961, C. acuminatum Vuxanovici, 1962, and C. kleini Foissner, 1969. The monotypic genus Dexiostoma contains D. campyla (Stokes, 1886) and is distinguished from Colpidium by a different oral infraciliature and the position and shape of the preoral suture. A new genus, Paracolpidium nov. gen., is suggested for Colpidium truncatum Stokes, 1885 because its oral infraciliature deviates from both Colpidium and Dexiostoma. Contrary to Colpidium and Dexiostoma, the silverline system of Paracolpidium truncatum lacks secondary meridians but shows short projections to the left of the primary meridians indicating a phylogenetic relationship to Tetrahymena.     

The ability of rumen ciliates, Eudiplodinium maggii, Diploplastron affine, and Entodinium caudatum, to use the murein saccharides

Murein polysaccharides may contribute to a considerable part of the dry matter of bacterial cells. Their utilization by protozoa inhabiting the rumen is, however, poorly recognized. The objective of this study was to examine the ability of three species of ciliates, i.e., Eudiplodinium maggii, Diploplastron affine, and Entodinium caudatum of digest, and ferment these saccharides. The cultivation experiments showed that the enrichment of growth medium with bacterial cell wall β-glycans increased the ciliate number (p?<?0.05). A statistically significant increase (p?<?0.01) was followed by a continuous decrease (p?<?0.01) in the percentage of individuals containing β-glycans particles after 4- and 24-h incubation of ciliates with this substrate, respectively. The enzymatic experiments confirmed the ability of the examined protozoa to digest murein. E. caudatum exhibited the highest activity (8.2 unit (U)/mg protein per min), and E. maggii, the lowest (3.0 U/mg protein per min). The production rates of volatile fatty acids by starved and fed ciliate species were 0.7 and 1.6 (E. caudatum)?pmol/ciliate cell per h, 30.5 and 42.5 (E. maggii)?pmol/ciliate cell per h, and 8.3 and 19.2 (D. affine)?pmol/ciliate cell per h (p?<?0.05).     

Cues from the predator crab Thalamita danae fed different prey can affect scope for growth in the prey mussel Perna viridis

Clearance rate (CR), absorption efficiency (AE), respiration rate, excretion rate and scope for growth (SFG) were investigated in the green-lipped mussel Perna viridis upon exposure to predatory crabs, Thalamita danae, that had been either starved or maintained on diets with P. viridis or shrimp tissue. The CR and SFG were significantly lower when the mussels were exposed to starved T. danae or those fed with mussels. The differences were observed immediately after the mussels were exposed to the cues (Day 0) and 7 days later. The AE, however, was significantly different among treatments on Day 0, with the highest efficiency being obtained for mussels exposed to crabs maintained on shrimp and followed by the control without any predator cues. Results showed that P. viridis was able to discriminate between predators on different diets and adjust physiological responses according to the level of perceived risk, with growth reduced at higher risks.     

Tolerance of a ruminant ciliate <Emphasis Type="Italic">Entodinium caudatum</Emphasis> against mercury,copper and chromium

The effects of three heavy metal cations, mercury (II), copper (II), and chromium (VI), on the growth of the rumen ciliate Entodinium caudatum in vitro culture was studied. The E. caudatum culture was challenged by HgCl₂, CuCl₂, and K₂Cr₂O₇ for a period of 4 days. The tested concentrations of mercury (II) and copper (II) were 1, 5, 10, 20, 50 mg/L and 2, 10, 20, 40 mg/L for chromium (VI) at single dose with either untreated or inhibited bacterial co-culture population. Effective metal concentrations required to reduce ciliate growth by 50% (EC₅₀) for mercury (II), copper (II), and chromium (VI) either with untreated or inhibited bacterial co-culture population after 24 h of metal application were 24, 20, and 21 or 15, 20, and 19 mg/L, respectively. After 4 days of metal application, corresponding EC₅₀ values for mercury (II), copper (II), and chromium (VI) were 16, 20, and 17 (with untreated bacterial population) or not determinable, 20, and 15 mg/L, respectively (with inhibited bacterial population). Increased sensitivity of E. caudatum to tested heavy metals with inhibited bacterial co-culture population indicate that the ciliate resistance to the heavy metal tested depends on detoxification abilities of rumen bacterial population.     

Nocodazole inhibits macronuclear infection with Holospora obtusa in Paramecium caudatum

Summary. Holospora obtusa is a Gram-negative bacterium inhabiting the macronucleus of the ciliate Paramecium caudatum. Experimental infection with H. obtusa was carried out under nocodazole treatment. Nocodazole has been shown to cause disassembly of the cytoplasmic microtubules radiating from the cytopharynx and postoral fibers in P. caudatum. Treatment with this drug did not prevent the ingestion of both prey bacteria and H. obtusa, but it reduced the phagosome number and affected cyclosis. In situ hybridization revealed infectious forms of this endobiont very close to the macronucleus, but never inside it. These results indicate that disassembly of microtubules does not impair transportation of the infectious forms of H. obtusa in the cytoplasm, but that it completely blocks the invasion of the nucleus by the bacteria. Correspondence and reprints: Department of Cytology and Histology, Faculty of Biology and Soil Sciences, Saint Petersburg State University, Universitetskaya naberezhnaya 7/9, 199034 Saint Petersburg, Russia.     

Predation on the invasive mosquito Aedes japonicus (Diptera: Culicidae) by native copepod species in Germany

Some limnic copepod species are predators of mosquito larvae. Seven species belonging to the order Cyclopoida, family Cyclopidae, were collected in the field in Germany and tested for the first time in laboratory bioassays for their potential to serve as biological control agents of the invasive Asian bush mosquito Aedes japonicus (Theobald), a vector of various pathogens causing disease. Females of Diacyclops bicuspidatus (Claus) did not attack 1^st instar larvae of Ae. japonicus, but Macrocyclops distinctus (Richard), Cyclops divergens Lindberg, and C. heberti Einsle predated a mean of 14, 18, or 19 1^st instar larvae, respectively. Acanthocyclops einslei Mirabdullayev and Defaye killed 30 larvae, and high predation rates with a mean of 39 or 46 larvae, respectively, were obtained by Megacyclops viridis (Jurine) and M. gigas (Claus). In regression analyses, predation rates by M. viridis correlated with body size, with specimens of 1.8 mm length being more effective than smaller or bigger ones. Based on the presented data, the two Megacyclops species seem to be promising candidates for use in field studies on the biological control of Ae. japonicus.     

Nuclear genes encoding chloroplast hemoglobins in the unicellular green alga Chlamydomonas eugametos

When the green unicellular alga Chlamydomonas eugametos is grown under light/dark regimes, nuclear genes are periodically activated in response to the changes in light conditions. These genetic responses are dependent upon the activation of genes associated with photosynthesis (LI616 and LI637), nonphotosynthetic photoreceptors (LI410 and LI818) and the biological clock (LI818). We report here that the LI410 and LI637 genes are part of a small gene family encoding hemoglobins (Hbs) related to those from two unicellular eukaryotes, the ciliated protozoa Paramecium caudatum and Tetrahymena pyriformis, and from the cyanobacterium Nostoc commune. Investigations of the intracellular localization of C. eugametos Hbs by means of immunogold electron microscopy indicate that these proteins are predominantly located in the chloroplast, particularly in the pyrenoid and the thylakoid region. To our knowledge, this constitutes the first evidence for the presence of Hbs in chloroplasts. Alignment of the LI637 cDNA nucleotide sequence with its corresponding genomic sequence indicates that the L1637 gene contains three introns, the positions of which are compared with those in the Hb genes of plants, animals and the ciliate P. caudatum. Although the LI637 gene possesses a three-intron/four-exon pattern similar to that of plant leghemoglobin genes, introns are inserted at different positions. Similarly the position of the single intron in the P. caudatum gene differs from the intron sites in the LI637 gene. The latter observations argue against the current view that all eukaryotic Hbs have evolved from a common ancestor having a gene structure identical to that of plant or animal Hbs.     

AM真菌影响入侵植物黄顶菊与本土物种狗尾草竞争生长的机理研究

该研究以入侵植物黄顶菊[Flaveria bidentis(L.)Kunt]和本土伴生植物狗尾草为材料,通过筛选出黄顶菊单一优势群落AM真菌,于温室盆栽条件下,采用2物种单播、混播以及接种AM真菌和不接种共6个处理,分析AM真菌对黄顶菊和狗尾草的根系侵染率、相对竞争强度、植株氮磷钾光合利用率、以及丙二醛含量和保护酶活性的影响,探讨AM真菌对黄顶菊与狗尾草竞争生长的机理。结果显示:(1)黄顶菊根际土壤AM真菌共包括4属10种,其中优势种为Glomus constrictum、Glomus perpusillum、Glomus reticulatum;盆栽接种AM真菌后,黄顶菊的根系侵染率显著高于本土伴生植物狗尾草,但接种AM真菌后黄顶菊相对竞争强度显著降低了29.57%,却对狗尾草相对竞争强度无显著影响。(2)接种AM真菌使黄顶菊植株光合氮、磷、钾利用率显著升高,但对伴生植物狗尾草的光合氮、磷、钾利用率均无显著影响。(3)接种AM真菌对黄顶菊植株POD和CAT活性以及MDA含量无显著影响,但显著增加了SOD和APX活性,而伴对生植物狗尾草的POD、CAT和APX活性均显著降低,MDA含量显著提高。研究表明,AM真菌对黄顶菊和狗尾草具有不同的选择性,AM真菌的定植促进了黄顶菊的竞争生长,增加了植株N、P含量、光合养分利用率以及抗氧化酶活性;但显著降低了本土伴生植物狗尾草的N、P吸收以及抗氧化酶活性。因此,AM真菌在竞争生长中对黄顶菊产生了偏利反馈,有助于黄顶菊的入侵。     

Promotion of Monacolin K Production by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-Mediated Transformation in <Emphasis Type="Italic">Monascus albidus</Emphasis> 9901

The binary vector pCAMBIA3300-gpdA-hph-trpC with hygromycin B phosphotransferase (hph) was constructed and transformed into Monascus albidus 9901 by Agrobacterium tumefaciens-mediated transformation, with gene hph as the selective marker. In order to improve the efficiency of A. tumefaciens-mediated transformation in M. albidus 9901, we optimized various factors including concentration of M. albidus 9901 spores, cell density of A. tumefaciens, co-cultivation time, temperature, and acetosyringone concentration. Most transformants of M. albidus 9901 could grow stably on media containing 50 μg ml⁻¹ hygromycin B up to five generations. The presence of hph was identified by PCR. Two transformants H1 and H2 which produced more Monacolin K than M. albidus 9901 were screened, and the concentration of Monacolin K in the fermented millet by H1 and H2 increased by 42.15% and 40.34% respectively compared with that produced by M. albidus 9901.     

The value of three cereal aphid species as food for a generalist predator

The value of the cereal aphid species Metopolophium dirhodum (Wlk.), Sitobion avenae (F.) and Rhopalosiphum padi (L.) as prey for the linyphiid spider Erigone atra (Bl.) was assessed. Fecundity of females was determined for spiders fed on eight experimental diets: three single‐species aphid diets, a mixed diet of all three aphid species, three mixed diets with each aphid species in combination with fruit flies Drosophila melanogaster (Meig.), and pure D. melanogaster as a high quality comparison diet. The development and survival of first‐instar juveniles fed on three diets of single aphid species, and on a diet of Collembola were compared with those subjected to starvation. Prey value for adult females was assessed by egg production, hatching success and offspring size. In pure diets all three aphid species were of low value to the spiders, causing a rapid decline in egg production and supporting no growth of significance of first‐instar juveniles. No difference in value of aphid species of single‐species aphid diets was found in the fecundity experiment, while a ranking of aphid species of M. dirhodum > R. padi > S. avenae was revealed in the survivorship experiment. A mixed‐aphid diet was not found to be advantageous compared with single‐species aphid diets, and no advantage of including aphids in mixed diets with fruit flies was found. Metopolophium dirhodum and R. padi were neutral in mixed diets, while a diet of S. avenae and fruit flies caused reduced egg production compared with the pure diet of fruit flies, revealing a toxic effect of S. avenae on the spider. The value‐ranking of aphid species in mixed diets was similar to that of single‐species diets. A similar ranking of aphid species was found for different fitness parameters (fecundity of adult females and development of juveniles). A ranking of aphids by offspring size of mothers on aphid‐only diets was S. avenae > M. dirhodum > R. padi. All aphid‐fruit fly diets resulted in larger offspring than a diet of only D. melanogaster, with the overall largest offspring being produced on the diet of M. dirhodum and fruit flies.     

Individual Specialization to Non-Optimal Hosts in a Polyphagous Marine Invertebrate Herbivore

Factors determining the degree of dietary generalism versus specialism are central in ecology. Species that are generalists at the population level may in fact be composed of specialized individuals. The optimal diet theory assumes that individuals choose diets that maximize fitness, and individual specialization may occur if individuals'' ability to locate, recognize, and handle different food types differ. We investigate if individuals of the marine herbivorous slug Elysia viridis, which co-occur at different densities on several green macroalgal species in the field, are specialized to different algal hosts. Individual slugs were collected from three original algal host species (Cladophora sericea, Cladophora rupestris and Codium fragile) in the field, and short-term habitat choice and consumption, as well as long-term growth (proxy for fitness), on four algal diet species (the original algal host species and Chaetomorpha melagonium) were studied in laboratory experiments. Nutritional (protein, nitrogen, and carbon content) and morphological (dry weight, and cell/utricle volume) algal traits were also measured to investigate if they correlated with the growth value of the different algal diets. E. viridis individuals tended to choose and consume algal species that were similar to their original algal host. Long-term growth of E. viridis, however, was mostly independent of original algal host, as all individuals reached a larger size on the non-host C. melagonium. E. viridis growth was positively correlated to algal cell/utricle volume but not to any of the other measured algal traits. Because E. viridis feeds by piercing individual algal cells, the results indicate that slugs may receive more cytoplasm, and thus more energy per unit time, on algal species with large cells/utricles. We conclude that E. viridis individuals are specialized on different hosts, but host choice in natural E. viridis populations is not determined by the energetic value of seaweed diets as predicted by the ODT.     

Survival of ciliate protozoa under starvation conditions and at low bacterial levels

Under starvation conditions, 50% survivorship times displayed no significant relationship with cell size in 2 ciliate species in this study and 5 protozoan species from the literature. Differences in survival ability were attributed to differences in weight-specific respiratory rate and relative motility among these 7 species. At low bacterial levels, 4 ciliate species in this study displayed significant differences in survivorship. High survivorship ofEuplotes patella relative to that ofParamecium caudatum andParaurostyla sp. at low ciliate densities was attributed to the lower individual energy requirements of this smaller species. High survivorship ofStentor coeruleus was interpreted as an effect of its large quantity of reserves and low respiratory rate. The survivorship ofE. patella was reduced at a higher population density. Four ciliate species survived longer at 15C than at 22C. Q₁₀ values based on 50% survivorship times at these 2 temperatures were much lower than Q₁₀ values based on respiratory rates and growth rates of well-fed ciliates over a similar temperature range.