Biotic factors in induced defence revisited: cell aggregate formation in the toxic cyanobacterium <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> PCC 7806 is triggered by spent <Emphasis Type="Italic">Daphnia</Emphasis> medium and disrupted cells

Bioassays with the toxic cyanobacterium Microcystis aeruginosa PCC 7806, its non-toxic mutant ΔmcyB, and Daphnia magna as grazer were used to evaluate biotic factors in induced defence, in particular cyanobacterial and grazer-released info-chemicals. Three main questions were addressed in this study: Does Daphnia grazing lead to a loss of cyanobaterial biomass? Is the survival time of Daphnia shorter in a culture of the toxic cyanobacterium? Does direct grazing or the presence of spent Daphnia medium or a high number of disrupted toxic Microcystis cells in the assays lead to an increase in the cellular microcystin content in the remaining intact cells? The biovolume (growth) as well as size and abundance of Microcystis aggregates were determined by particle analysis, while the survival time of Daphnia individuals was recorded by daily observation and counting, with the relative concentration of cell-bound microcystin-LR, was measured by HPLC analysis. Compared to some recent studies in the field of induced defence, in this study, evidence was found for a direct grazing effect, i.e. the loss of biovolume in the toxic culture. In addition, Daphnia magna ingested more non-toxic than toxic cells, and survived longer with non-toxic cells. In terms of increased cell-bound toxin concentration as a means of defence reported in some studies, a higher cell-bound microcystin-LR content was not measured in this study in any of the treatments (P > 0.05). Under low light conditions with impaired growth of Microcystis, and the presence of a high number of particles with less than 1-μm diameter (possibly heterotrophic bacteria), Daphnia medium was associated with a strong reduction in cell-bound toxin concentration (P < 0.05). This study showed no increased cell aggregation under direct grazing (P > 0.05), but increased aggregation with spent Daphnia medium under high light conditions (P < 0.05). Further, the addition of cell-free extract from disrupted toxic Microcystis cells strongly increased the aggregation of the intact cells under low light (P < 0.05). These findings are discussed with the possible role of microcystin and other infochemicals in the expression of proteins and morphology changes in Microcystis.     

Freshwater anostracan, <Emphasis Type="Italic">Branchinella kugenumaensis</Emphasis>, as a potential controlling consumer species on toxic cyanobacteria <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

To evaluate the potential of Branchinella kugenumaensis for cyanobacterial bloom control relative to Daphnia, we conducted several feeding experiments on microcystin-free and microcystin-containing unicellular strains of Microcystis aeruginosa and colonial forms of Microcystis using B. kugenumaensis and Daphnia magna in a laboratory. Branchinella kugenumaensis showed higher filtration rates than those of D. magna in all treatments. In particular, the microcystin-containing unicellular strain supported the highest filtration rates of B. kugenumaensis among treatments. Daphnia magna reduced colonies less than 75 μm in length, whereas B. kugenumaensis could graze colonies less than 100 μm. The middle-sized group of B. kugenumaensis had a higher filtration rate than the small and large sized groups in a continuous feeding experiment for 4 days. In survival experiments, survivorships were not different between the two species, whereas ages at the beginning of the experiments affected their survival time. Our results showed that B. kugenumaensis grazed on toxic and colonial cyanobacteria at relatively high rates, indicating that locally abundant grazers like Branchinella may offer a better potential for bloom control than Daphnia.     

Effect of the vital activity products of beaver (<Emphasis Type="Italic">Castor fiber</Emphasis> L.) on the formation of zooplankton structure: Changes in the quantitative parameters of two cladocera species of different sizes in a beaver pound (in situ experiment)

The experiments were performed in microcosms situated on a control site and on a site regulated by beavers. Two cladoceran species of different sizes were placed in microcosms in different proportions. It was found that the vital activity products of Castor fiber L. promote an increase in the concentration of total phosphorus (P), a decrease in the N/P ratio in water, and an increase in the chlorophyll a concentration and the abundance and biomass of bacterioplankton. In such conditions, the abundance and biomass of the large species Daphnia (Ctenodaphnia) magna Straus was increased to the greatest extent when compared with background values when it was placed in combination with Ceriodaphnia dubia Richard or without it. Moreover, in the case of the combination of two crustacean species in microcosms, the abundance of C. dubia was lower when compared with the control site, while the abundance of D. magna was higher. The results of a bioassay showed that the productivity of C. dubia decreases in waters where D. magna is most abundant. We concluded that the formation of zooplankton in beaver ponds with specific features determined by the presence of large cladoceran species is provided by changes in the quantitative and qualitative parameters of food resources. This contributes to the massive distribution of large D. magna and its competitive relations with small C. dubia. According to the results of a bioassay, the vital activity products of D. magna can inhibit the fecundity of C. dubia.     

To eat or not to eat infected food: a bug’s dilemma

Parasites can adversely affect host population densities, but predators can regulate disease by reducing the density of susceptible hosts and consuming parasites contained in infected hosts. Some parasites induce phenotypic modifications in their hosts that potentially lead to increased predation. We investigated the role of parasite-induced modified appearance in the interactions between the crustacean Daphnia magna, its bacterial parasite Pasteuria ramosa, and its predator, the backswimmer Anisops sp. Our aim was to test the backswimmer’s prey preference between infected and uninfected D. magna to find out whether infection by P. ramosa can affect predation risk by Anisops. We found that Anisops sp. had a strong preference for uninfected D. magna under light, but under dark conditions the preference was reversed, which suggests that the modified appearance is the cause of this preference. Such anti-parasite preference by Anisops sp. could strongly influence host population dynamics as loss of fecundity, disease mortality, and predation are additive, resulting in host population decline.     

The Plankton Community of Sevan Lake (Armenia) after Invasion of <Emphasis Type="Italic">Daphnia</Emphasis> (<Emphasis Type="Italic">Ctenodaphnia</Emphasis>) <Emphasis Type="Italic">magna</Emphasis> Straus, 1820

The appearance of Daphnia (Ctenodaphnia) magna Straus, 1820 in the pelagial of Sevan Lake caused significant changes in the communities of planktonic algae, bacteria, and heterotrophic nanoflagellates. Phytoplankton and nanoflagellates were the most affected by the direct impact of D. magna, and the number and biomass of bacteria increased due to the reduction in the trophic pressure of the protists. It was also facilitated by the increased supply of phosphorus as a result of the activity of the cladocerans, as well as the decrease in the number and biomass of phytoplankton, which competes with heterotrophic bacteria for the nutrients.     

Genetic polymorphism,haplotype distribution,and phylogeny of <Emphasis Type="Italic">Daphnia</Emphasis> (Cladocera: Anomopoda) species from the water bodies of russia as inferred from the 16S mtDNA gene sequencing

The data on the genetic polymorphism of the most widespread Daphnia species occupying different water bodies of Russia are presented. The phylogenetic relationships between the examined species were established, and the haplotype networks were constructed. A fragment of the 16S mitochondrial DNA gene was used as a genetic marker. The results of molecular phylogenetic analysis generally coincided with modern concepts in the systematics of the genus Daphnia. The representatives of the divergent mitochondrial lineages within the D. longispina, D. pulex, and D. magna complex remain poorly investigated morphologically. For D. dentifera, a new habitat on the territory of Russia, namely, the water bodies of the Lake Baikal basin, was identified. A conclusion was made that the 16S mtDNA gene could be successfully used in phylogeographic analysis of the genus Daphnia.     

Experimental studies of the effect of beaver (<Emphasis Type="Italic">Castor fiber</Emphasis> L.) vital activity products on the formation of zooplankton structure (by the example of growth of two cladoceran species of different sizes)

Experiments in microcosms have demonstrated that beaver vital activity products (BVAPs) promote an increase in concentrations of total nitrogen (N) and total phosphorus (P), a decrease in the N/P value in water, and an increase in the abundance and biomass of bacterioplankton. Under such conditions, the abundance and biomass of small Ceriodaphnia dubia Richard and large Daphnia (Ctenodaphnia) magna Straus, which live separately, increase. The coexistence of these cladocerans in microcosms under the BVAP influences results in a high increase in the abundance and biomass of D. magna; in similar experiments without the influence of BVAP, Ceriodaphnia dubia becomes more abundant. The results of bioassay demonstrate that the number of newborns of Ceriodaphnia dubia decreases in water where Daphnia magna is numerous owing to BVAPs. It is suggested that the vital activity products of large representatives of the genus Daphnia inhibit the fecundity of small species of Cladocera. This fact, along with the high competitiveness of large cladoceran species under conditions of a high level of nutritive base, determine the formation of zooplankton communities in beaver ponds which are characterized by a high abundance and biomass and low uniformity     

Dynamic variation of toxic and non-toxic <Emphasis Type="Italic">Microcystis</Emphasis> proportion in the eutrophic Daechung Reservoir in Korea

This study was conducted to determine the environmental factors affecting the level of potentially toxic Microcystis. The long-term tendencies of temperature, precipitation, and water quality factors were analyzed to determine the environmental characteristics of the Daechung Reservoir in Korea, and water samples were directly collected to analyze the dynamics of toxic and non-toxic Microcystis at weekly intervals from May to October 2012. Microcystis was the dominant genus during the study period, and it was composed of potentially toxic and non-toxic Microcystis. The fraction of potentially toxic Microcystis ranged from 6.0% to 61.1%. The amount of toxic Microcystis was highly related to the intracellular microcystin concentration (r = 0.760, P < 0.01). Therefore, the fraction of potentially toxic Microcystis is an important concern in Microcystis blooming because the intracellular microcystin concentration may reflect microcystin levels in the water. The prevalence of potentially toxic Microcystis was highly related to water temperature in Daechung Reservoir (r = 0.585, P < 0.01). Thus, temperature increase during Microcystis blooming may lead to more frequent toxic Microcystis blooms in eutrophic water bodies.     

Acute toxicity of copper to <Emphasis Type="Italic">Daphnia galeata</Emphasis> under different magnesium and calcium conditions

A number of risk assessments of the adverse effects of Cu and its compounds have been conducted since it is one of the leading substances responsible for water contamination. However, with the exception of standard organisms, ecotoxicological knowledge is still scarce. Here, we examined the influence of Ca²⁺ and Mg²⁺ concentrations on acute toxicity of Cu toward a cladoceran, Daphnia galeata Sars. We found that the protective effects of Ca²⁺ against copper toxicity were larger than those of Mg²⁺. BLM parameters (log Ks) for Ca and Mg estimated based on our 48-h LC₅₀ (50 % lethal concentration), respectively, were 3.14 and 2.29. The log K of Ca was similar to that reported in previous studies using Daphnia magna; however, the log K of Mg was lower by one order of magnitude. Our results suggest that there is some mechanistic difference related to the Mg uptake between D. magna and D. galeata. The results obtained from the present study will contribute to the water quality criteria of copper in soft water because D. galeata is a widespread (distributed in Eurasia and North America) and common species, even in soft water lakes.     

Toxin Gene Contents and Activity of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Strains Against Two Sugarcane Borer Species, <Emphasis Type="Italic">Diatraea saccharalis</Emphasis> (F.) and <Emphasis Type="Italic">D. flavipennella</Emphasis> (Box)

Bacillus thuringiensis (Berliner) bears essential characteristics in the control of insect pests, such as its unique mode of action, which confers specificity and selectivity. This study assessed cry gene contents from Bt strains and their entomotoxicity against Diatraea saccharalis (F.) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). Bioassays with Bt strains were performed against neonates to evaluate their lethal and sublethal activities and were further analyzed by PCR, using primers to identify toxin genes. For D. saccharalis and D. flavipennella, 16 and 18 strains showed over 30% larval mortality in the 7th day, respectively. The LC₅₀ values of strains for D. saccharalis varied from 0.08 × 10⁵ (LIIT-0105) to 4104 × 10⁵ (LIIT-2707) spores + crystals mL^?1. For D. flavipennella, the LC₅₀ values of strains varied from 0.40 × 10⁵ (LIIT-2707) to 542 × 10⁵ (LIIT-2109) spores + crystals mL^?1. For the LIIT-0105 strain, which was the most toxic to D. saccharalis, the genes cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, cry8, and cry9C were detected, whereas for the strain LIIT-2707, which was the most toxic to D. flavipennella, detected genes were cry1Aa, cry1Ab, cry1Ac, cry1B, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, and cry9. The toxicity data and toxin gene content in these strains of Bt suggest a great variability of activity with potential to be used in the development of novel biopesticides or as source of resistance genes that can be expressed in plants to control pests.     

Isolation and Characterization of Toxic Cyanobacteria from Different Natural Sources

We isolated seven algologically and five bacteriologically pure cultures of toxin-producing cyanobacteria from Turgen gorge (Kazakhstan), Karlovy Vary (Czech Republic), and Shar-Nuur Lake, Bayan Ulgiiregion (Mongolia) springs. According to the Daphnia magna test, Desertifilum sp. and Nostoc sp. strains were the most toxic in the test of isolated strains (complete death of all test organisms was detected after 48 h). These strains possessed the highest inhibitory effect on proliferation of the HeLa cancer cell line. The Anabaena sp. 35 and Nostoc sp. 4 strains were also high toxic. Model strains Synechocystis PCC 6803 and Synechococcus elongates PCC 7942, as well as the strain isolated in the present work, Synechococcus sp. 55, were less toxic. Mass spectrometry made it possible to assign cyanobacterial toxins to cyclic depsipeptides. Two cyclic depsipeptides, micropeptin T and oscillapeptin, were detected in Desertifilum sp. extracts. Cryptophycin and small amounts of cyclic depsipeptide micropeptin SD were detected in Nostoc sp. extract.     

Host Foraging Behavior of <Emphasis Type="Italic">Dastarcus helophoroides</Emphasis> (Coleoptera: Bothrideridae) Adults,a Coleopteran Parasitoid of <Emphasis Type="Italic">Monochamus alternatus</Emphasis> (Coleoptera: Cerambycidae)

Dastarcus helophoroides adults, a coleopteran parasitoid of Monochamus alternatus, were reared in the laboratory. The objectives of this paper are to investigate the host foraging behavior of D. helophoroides, and the evidence for learning in such behaviors. Choice and no-choice experiments were carried out. The choice experiments show that the number of D. helophoroides adults choosing the Pinus massoniana log infested with M. alternatus larvae was significantly higher than that choosing the blank (a flavorless simulative log made of brown kraftpaper), and the number of D. helophoroides adults choosing the uninfested P. massoniana log was also significantly higher than that choosing the blank. The no-choice experiment shows no significant difference between the number of D. helophoroides adults that chose the infested log and that chose the uninfested log, despite previous experience with uninfested log, infested log, or neither of them. No evidence of learning in host foraging behavior in D. helophoroides adults was found. The results indicate that the volatile chemicals emitted by uninfested P. massoniana logs and infested logs with M. alternatus larvae are equally attractive to D. helophoroides.     

Trail Chemicals of the Convergens Ladybird Beetle, <Emphasis Type="Italic">Hippodamia convergens</Emphasis>, Reduce Feeding and Oviposition by <Emphasis Type="Italic">Diaphorina citri</Emphasis> (Hemiptera: Psyllidae) on Citrus Plants

We investigated feeding and oviposition behavior of the Asian citrus psyllid, Diaphorina citri, when exposed to the foraging trails of the convergens ladybird beetle, Hippodamia convergens. Diaphorina citri females feeding on citrus leaves directly exposed to the ladybird adults or treated with trail extract excreted significantly less honeydew droplets than controls. The trail chemicals of the ladybird beetle also decreased oviposition by D. citri females on citrus. In a no-choice experiment, D. citri females preferred to oviposit on control flush and plants than those with ladybird trail-extract treatments. In two-choice experiments, 68.0% of D. citri released into cages exhibited strong selection preference for settling and eventual oviposition on control plants than plants treated with ladybird trail extract. Diaphorina citri eggs were found on all new leaf flush of control plants, whereas only 29.5% of flush on treatment plants were selected for oviposition. The trail chemical deposited by the convergens ladybird beetle elicits repellency of D. citri feeding and oviposition. Therefore, the trail chemicals my contain components that could be useful for behavior-based management of D. citri and HLB disease by reducing psyllid feeding and oviposition.     

Assessment of atrazine decontamination by epiphytic root bacteria isolated from emergent hydrophytes

Aim of the study was to identify atrazine remediating bacteria that can potentially succeed in situ where they encounter varied environmental conditions. Three epiphytic root bacteria, genus Pseudomonas and Arthrobacter, were isolated from rhizoplanes of hydrophytes Acorus calamus, Typha latifolia, and Phragmites karka. Potential of these strains to decontaminate environmentally relevant concentrations of atrazine was determined in liquid atrazine medium (LAM) and Luria-Bertani (LB) medium at varying pH and temperature. There was an increase in decontamination by the strains with time upon exposure to 2.5 to 10 mg l^?1 atrazine over a period of 15 days, notably, in both minimal and nutrient-rich media. Growth in terms of O.D.₆₀₀ and biomass determined during the same period also showed a corresponding surge. Pseudomonas sp. strain AACB mitigated atrazine in a wide range of pH (5 to 8). Pseudomonas sp. strains AACB and TTLB decontaminated >?62% atrazine at 10 °C. All the strains exhibited plant growth–promoting traits in vitro, reported for the first time in the presence of atrazine. Strain AACB exhibits the novel trait of atrazine decontamination under harsh environmental conditions mimicked in lab. Strains isolated in the present study promise success in in situ remediation. Bioreactors and water treatment plants can be designed comprising the hydrophytes and the strains inoculated into their rhizospheres to improve efficacy of the treatment. They can be used to study plant-bacterium mutualistic symbiosis or other interactions occurring during atrazine mitigation.     

Novel rhodanine derivatives are selective algicides against <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

In this study, a series of rhodanine derivatives containing various substituents was synthesized and tested for in vitro algicidal activity. Among the tested substituent groups, phenyl substituents with halogen groups showed good inhibitory potency. Furthermore, the compound with chlorine at the C2 position of the phenyl ring exhibited a higher algicidal effect than the compound with chlorine at the C3 position of the phenyl ring. Among the various rhodanine derivatives tested, 5-(2,4-dichlorobenzylidene)- rhodanine (compound 20) was the most potent inhibitor against M. aeruginosa with a lethal concentration 50 (LC₅₀) value of 0.65 μM and Selenastrum capricornutum with an LC₅₀ value of 0.82 μM. To verify the feasibility of their use in ecosystems, 25 h of acute ecotoxicity tests were carried out for three derivatives against Danio rerio and Daphnia magna. No mortality was observed in groups exposed to 2.0 μM of compound 20 after 100 h of exposure. Moreover, a survival rate of 100% was observed in D. magna exposed to 2 μM of compound 20 for 100 h. Overall, the results show that rhodanine derivatives are a possible method for controlling and inhibiting harmful algal blooms.     

Revealing physiological and genetic properties of a dominant maize dwarf <Emphasis Type="Italic">Dwarf11</Emphasis> (<Emphasis Type="Italic">D11</Emphasis>) by integrative analysis

Plant height is an important agronomic trait involved in lodging resistance and harvest index. The identification and characterization of mutants that are defective in plant height have implications for trait improvement in breeding programs. Two dominant maize dwarf mutants D8 and D9 have been well-characterized. Here, we report the characterization of a dominant maize dwarf mutant Dwarf11 (D11). Dwarf stature of D11 was mainly attributed to the inhibition of longitudinal cell elongation. The levels of bioactive GA₃ were significantly lower in D11. Contrarily, D8 mutant accumulates markedly higher levels of GA₃. The expression of GA biosynthetic and catabolic genes was dramatically decreased in D11. Expression variations of d8 and d9 genes were not observed in D11 mutant. Moreover, genetic suppressors of D11 were identified in inbred line Chang 7-2. Integrated omics data indicated that D11 is a novel dominant maize dwarf. The ultimate D11 gene cloning and its regulatory network elucidation may strengthen our understanding of the genetic basis of plant architecture and provide cues for breeding of crops with plant height ideotypes.     

Contribution of olfactory and gustatory sensations of octanoic acid in the oviposition behavior of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> (Diptera: Drosophilidae)

Selection of oviposition sites in insects represents an important part of their ecological adaptation. In Drosophila fruit flies, adult preference for a particular oviposition site determines larval food, affecting fitness throughout the entire life cycle. Two odorant-binding proteins (OBPs) OBP57d and OBP57e were identified to be involved in the evolution of specific preference for the toxic plant Morinda citrifolia L. in D. sechellia Tsacas &; Bächli. D. melanogaster Meigen mutants for Obp57d and Obp57e showed enhanced preference for octanoic acid, but still not as much as D. sechellia does, indicating that other genes are also involved in the behavioral evolution of D. sechellia. Here, by using an improved method for behavioral assay, we found that the ablation of antenna enhanced the preference for octanoic acid in the Obp57d and Obp57e mutants to a level comparable with D. sechellia, suggesting that both olfactory and gustatory sensations are involved in oviposition site selection in response to octanoic acid. Behavioral analysis of gene-knockout strains revealed that Odorant receptor co-receptor (Orco) has little contribution compared with the effect of antennal ablation. These data suggest that in addition to Obp57d and Obp57e, the evolution of D. sechellia involves genetic changes in olfactory genes that function independently from Orco.     

A comparative cytogenetic study of the tetraploid oat species with the A and C genomes: <Emphasis Type="Italic">Avena insularis,A. magna</Emphasis>, and <Emphasis Type="Italic">A. murphyi</Emphasis>

C-banding of chromosomes and in situ hybridization with the probes pTa71 and pTa794 were used for a comparative cytogenetic study of the three tetraploid oat species with the A and C genomes: Avena insularis, A. magna, and A. murphyi. These species were similar in the structure and C-banding patterns of several chromosomes as well as in the location of the loci 5S rRNA genes and major NOR sites; however, they differed in the number and localization of minor 45S rDNA loci as well as in the morphology and distribution of heterochromatin in some chromosomes. According to the data obtained, A. insularis is closer to A. magna, whereas A. murphyi is somewhat separated from these two species. Presumably, all the three studied species originated from the same tetraploid ancestor, and their divergence is connected with various species-specific chromosome rearrangements. The evolution of A. murphyi is likely to have occurred independently of the other two species.     

Seasonal detection of atrazine and <Emphasis Type="Italic">atzA</Emphasis> in man-made waterways receiving agricultural runoff in a subtropical,semi-arid environment (Hidalgo County,Texas, USA)

Atrazine is a widely-used herbicide that can impact non-target organisms in the environment but can be biologically degraded by several types of microorganisms. In this study, the gene atzA, which encodes for the initial step in bacterially-mediated atrazine degradation, was used as an indicator of atrazine pollution in agricultural canals located in Hidalgo County, Texas, USA. The concentration of atrazine and atzA were monitored once per month for 12 months during 2010–2011. Atrazine was measured using an enzyme-linked immunosorbent assay; atzA abundance was monitored using Quantitative Polymerase Chain Reaction (Q-PCR) analyses. Abundance of atrazine and atzA were compared with rainy versus dry months and during planting versus non-planting months. Results showed that atrazine levels varied from below detection to 0.43 ppb and were not influenced by precipitation or planting season. Concentrations of the gene atzA were significantly different in rainy versus dry months; during planting versus non-planting times of the year; and in the interaction of precipitation and planting season. The highest concentration of atzA, approx. 4.57?×?10⁸ gene copies ml^?1, was detected in July 2010—a rainy, planting month in Hidalgo County, South Texas. However, atrazine was below detection during that month. We conclude that Q-PCR using atzA as an indicator gene is a potential method for monitoring low levels of atrazine pollution in environmental samples.