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Glycosidases in Brachionus plicatilis (Rotifera).

1. Tests for glycosidases were performed in homogenates of Brachionus plicatilis. 2. Hydrolytic activity was detected with the following substrates: (a) with synthetic substrates (NP = 4-nitrophenyl): NP-alpha- and NP-beta-D-glucopyranoside, NP-alpha- and NP-beta-D-galactopyranoside, NP-N-acetyl-beta-D-glucosaminide, NP-N-acetyl-beta-D-galactosaminide, NP-alpha- and NP-beta-D-mannopyranoside and NP-alpha-L-fucopyranoside; (b) with disaccharides: sucrose, maltose, trehalose, isomaltose, cellobiose, gentiobiose and lactose; (c) with polysaccharides: laminarine, carboxymethyl-cellulose, avicel, Micrococcus luteus (for lysozyme) and 4-nitrophenyl-alpha-D-maltoheptaoside (for amylase). 3. The pH dependence of the glycosidase activities was determined. 4. The distribution of enzyme activities within fractions from the homogenate was studied in order to localize them within the cell. 5. Proteins from Brachionus homogenate were separated by SDS-gel electrophoresis and the positions of the following glycosidase activities were detected by assays performed on the gels (estimated molecular weights in parentheses): alpha-glucosidase (250,000); beta-glucosidase (200,000); beta-galactosidase (70,000); N-acetyl-beta-glucosaminidase (60,000).

Animals↗

Rotifer neuropharmacology--I. Cholinergic drug effects on oviposition of Philodina acuticornis (Rotifera, Aschelminthes).

The presence of acetylcholine in the culture medium was found to result in egg retention in the bdelloid rotifer Philodina acuticornis in a dose-dependent fashion, with no other discernible physiological effect. When six to eight eggs accumulate, the animals distend, burst and die. No other cholinergic agonist has been shown to produce a similar effect. The antagonistic effect of six anticholinergic drugs and five acetylcholinesterase-inhibitor insecticides was investigated on egg retention. All compounds were found to inhibit this phenomenon to varying degrees, but on the basis of their EC50 neuromuscular blockers appear to be most active in inhibiting egg retention. We suggest, therefore, that egg retention is caused by a spasm or increased tone of the cloacal sphincter rather than a paralysis of muscles needed in the peristaltic expulsion of the egg. Five acetylcholine-esterase inhibitory insecticides also inhibit egg retention.

Acetylcholine↗

Rotifer neuropharmacology--II. Synergistic effect of acetylcholine on local anesthetic activity in Brachionus calyciflorus (Rotifera, Aschelminthes).

A number of compounds showing general anesthetic action in the rotifer Brachionus calyciflorus were investigated in the presence of acetylcholine. Non-ionizing anesthetics, including tricaine, showed no interaction with acetylcholine. However, highly ionized compounds like the local anesthetics procaine and lidocaine, the muscarinic blocker and local anesthetic atropine, and the beta-adrenergic blocker propranolol showed a synergistic effect with acetylcholine. ACh increased the general anesthetic effect of these compounds in a statistically highly significant dose-dependent fashion. To account for the mechanism of this unusual and novel effect it is proposed that these compounds interact with the anesthetic binding site of the rotifer cholinoceptor ionophore in the open state. It is also proposed that non-ionizing compounds have a general membrane effect only. In addition to anesthesia, atropine and propranolol cause foot paralysis in B. calyciflorus. This other novel effect is also enhanced by acetylcholine as well as decamethonium, a neuromuscular blocker.

Acetylcholine↗

Combined effects of food concentration and the herbicide 2,4-dichlorophenoxyacetic acid on the population dynamics of Brachionus patulus (Rotifera).

Herbicides are important in crop protection and management. A number of them including 2,4-D (2,4-dichlorophenoxyacetic acid), however, may reach water bodies and eventually affect the non-target organisms such as rotifers. In the present work, we studied the influence of 6 concentrations viz. 0 (control), 100, 200, 300, 400 and 500 mg l-1 of 2,4-D on the population growth of the rotifer Brachionus patulus under two algal (Chlorella) food levels (0.5 x 10(6) and 1.5 x 10(6) cells ml-1). Regardless of herbicide concentration, the population growth of B. patulus was dependent on the algal food levels, in that an increase in algal food level supported a better population growth. Similarly at any Chlorella density, the herbicide had a negative influence on the population growth of B. patulus. Herbicide level of 500 mg l-1 inhibited population growth of B. patulus beyond 5 days. Rotifers grown under low food density and high herbicide concentration (300 mg l-1 or above) were completely eliminated after day 15. The rate of population increase (r) (mean +/- standard error) in the controls varied from 0.46 +/- 0.002 and 0.55 +/- 0.004 under 0.5 x 10(6) and 1.5 x 10(6) cells ml-1 of Chlorella, respectively. The r values became negative under both, low and high food levels, at or beyond 300 mg l-1 of 2,4-D. The maximal population abundance (ind. ml-1) in controls varied from 294 +/- 9 to 503 +/- 21 under low and high food levels of Chlorella, respectively. The role of algae in mitigating adverse effects of high herbicide concentrations to rotifers has been discussed.

2,4-Dichlorophenoxyacetic Acid↗

Effects of mercury on the life table demography of the rotifer Brachionus calyciflorus Pallas (Rotifera).

Mercury is highly toxic to a variety of aquatic organisms including zooplankton. The functioning of freshwater ecosystems can be altered if rotifers, being a natural food link between phytoplankton and fish larvae, are contaminated by mercuric compounds. In order to detect age-specific responses of the rotifer Brachionus calyciflorus to mercury toxicity (5 nominal concentrations as chloride viz. 0, 0.000625, 0.00125, 0.0025 and 0.005 mg l(-1)), we used the standard life table method at two different food (Chlorella vulgaris) levels (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1)). Data indicated that increase in mercury concentration had an increasingly intense negative effect on many of the life history variables, while at higher food levels, its impact was less. A nearly rectangular survivorship pattern was obtained in controls, especially at higher food levels. This trend gradually changed to a steep fall as the concentration of the heavy metal in the medium increased from 0 to 0.005 mg l(-1). At any given food density, increase in the mercury concentration resulted in decreased age-specific reproduction. A maximum of 3.5 offspring female(-1) was observed in controls at higher food density. The average lifespan varied from 6 to 8 days at low food level, depending on the heavy metal concentration in the medium. The corresponding values at high food level varied from 8 to 12 days. Regardless of mercury concentration in the medium, gross and net reproductive values varied from 10 to 33 and 4 to 19 offspring female(-1). The longest generation time (about 9 days) of B. calyciflorus was obtained at 1.5 x 10(6) cells ml(-1) food density in control, while the shortest was 5 days at low food level and high (0.005 mg l(-1)) mercury concentration in the medium. Depending on the food level and heavy metal concentration in the medium, the rate of population growth (r) varied from 0.32 to 0.62 d(-1). In general, higher food level resulted in higher r. Except generation time, all other derived variables were significantly influenced by food level and the heavy metal concentration in the medium.

Analysis of Variance↗

Divergent gene copies in the asexual class Bdelloidea (Rotifera) separated before the bdelloid radiation or within bdelloid families.

Rotifers of the asexual class Bdelloidea are unusual in possessing two or more divergent copies of every gene that has been examined. Phylogenetic analysis of the heat-shock gene hsp82 and the TATA-box-binding protein gene tbp in multiple bdelloid species suggested that for each gene, each copy belonged to one of two lineages that began to diverge before the bdelloid radiation. Such gene trees are consistent with the two lineages having descended from former alleles that began to diverge after meiotic segregation ceased or from subgenomes of an alloploid ancestor of the bdelloids. However, the original analyses of bdelloid gene-copy divergence used only a single outgroup species and were based on parsimony and neighbor joining. We have now used maximum likelihood and Bayesian inference methods and, for hsp82, multiple outgroups in an attempt to produce more robust gene trees. Here we report that the available data do not unambiguously discriminate between gene trees that root the origin of hsp82 and tbp copy divergence before the bdelloid radiation and those which indicate that the gene copies began to diverge within bdelloid families. The remarkable presence of multiple diverged gene copies in individual genomes is nevertheless consistent with the loss of sex in an ancient ancestor of bdelloids.

Animals↗

Effect of cadmium and chromium toxicity on the demography and population growth of Brachionus calyciflonrs and Brachionus patulus (Rotifera).

This paper reports on the assessment of the effect of Cd+2 and Cr+6 to the rotifers Brachionus calyciflorus and B. patulus using life table demography and population growth. Based on acute toxicity tests, cadmium was nearly 100 times more toxic than chromium for both the rotifer species. Age-specific fecundity curves of both the brachionids showed increased offspring beyond the age of 4 days and after that the production nearly stabilized for about 10 days and later declined. At any given treatment, B. calyciflorus had the higher rate of offspring production than B. patulus. For any given rotifer, increased metal concentration (Cd or Cr) resulted in decreased offspring production. While chromium at a concentration of 8.0 mg L(-1), permitted reproduction in B. calyciflorus, the egg production of B. patulus was inhibited at half that level (4.0 mg L(-1)). Cd at concentrations as low as 0.056 mg L(-1) caused 10-30% reduction in survivorship and reproductive parameters of B. calyciflorus. The lowest chromium concentration chosen for B. calyciflorus (0.5 mg L(-1)) had, however, a lower impact (2-22%) on the life history variables. Corresponding values of life history variables for B. patulus at the lowest Cd (0.0028 mg L(-1)) and Cr (0.25 mg L(-1)) concentrations varied greatly (2-40% and < 1-24%, respectively). Compared to controls, both the rotifer species in Cd or Cr treatments had lower densities. In controls, B. calyciflorus reached a peak abundance of about 100 ind. mL(-1), while the corresponding value for B. patulus was slightly higher (120 ind. mL(-1)). The rate of population increase (r) derived from the population growth data varied from +0.42 to -0.40 depending on the concentration of the heavy metal in the medium.

Animals↗

The combined effects of zinc and alga on the life table demography of Anuraeopsis fissa and Brachionus rubens (Rotifera).

In this work the combined effects of zinc and Chlorella on the demography of two co-occurring planktonic rotifers (A. fissa and B. rubens) were evaluated. Chlorella absorbed a significant (about 35%) quantity of zinc from the medium within the 24 h period. The age-specific survivorship curves of A. fissa and B. rubens showed increased mortality rate with increasing zinc concentration in the medium (0 to 0.5 mg L(-1)), at the two Chlorella levels (0.5 x 10(6) and 1.0 x 10(6) cells mL(-1)). In controls, both the rotifer species showed enhanced survival at higher food level (1.0 x 10(6) cells mL(-1)). Low food level (0.5 x 10(6) cells mL(-1)) and higher concentration (0.25 and 0.5 mg L(-1)) of zinc resulted in a steep fall in the survivorship. Regardless of Zn concentration and food level, A. fissa had a shorter average lifespan, lower gross and net reproductive rate, and rate of population increase than B. rubens. The maximum offspring production on a given day in controls for A. fissa was about 2 female(-1), which was reduced to half when exposed to 0.5 mg L(-1) of Zn. At both food levels, A. fissa exposed to Zn at 0.5 mg L(-1) had negative population growth rates. The highest population growth rate (0.94 day(-1)) was observed for B. rubens in controls at 1.0 x 10(6) cell mL(-1) of Chlorella. The present data showed that A. fissa was more sensitive to zinc toxicity than B. rubens. Combination of high vulnerability to metal toxicity and low population growth rate, A. fissa in zinc contaminated waterbodies could be possibly more adversely affected than B. rubens.

Animals↗

Effect of cadmium level and exposure time on the competition between zooplankton species Moina macrocopa (Cladocera) and Brachionus calyciflorus (Rotifera).

Competition among zooplankton is a natural phenomenon and often cladocerans are competitively superior to rotifers. However, anthropogenic factors including the release of industrial effluents, may influence this interaction. In this study, we evaluated the effect of cadmium (0.05 and 0.1 mg L(-1) as CdCl(2)) on competition between the cladoceran Moina macrocopa and the rotifer Brachionus calyciflorus. Since the release of industrial effluents is generally pulsed, we also exposed the test zooplankton species at different exposure periods (3, 6, 12 and 24 h) to the heavy metal. Regardless of exposure time and the presence of competing species, an increase in concentration of Cd resulted in decreased population growth of M. macrocopa and B. calyciflorus. Regardless of presence of the competing species and Cd concentration, an increase in exposure period resulted in decreased population growth rates of both the zooplankton species. In mixed cultures, in general, M. macrocopa outcompeted B. calyciflorus and completely eliminated it under conditions of high toxicant concentrations and longer exposure time.

Animals↗

Population growth of Euchlanis dilatata (Rotifera): combined effects of methyl parathion and food (Chlorella vulgaris).

In the present work, the combined impact of four concentrations (0, 0.0625, 0.125, and 0.25 mg/L) of methyl parathion and three densities (0.5 x 10(6), 1.0 x 10(6), and 2.0 x 10(6) cells/mL) of the green alga Chlorella vulgaris on the population growth of Euchlanis dilatata was studied. In general, regardless of the food level, an increase in the concentration of methyl parathion resulted in a significant reduction of the maximal population density and rate of population increase. The population growth rate in the controls ranged from 0.248 to 0.298; rates were lower in the presence of the pesticide. At any toxicant concentration, rotifers fed higher algal density showed significantly higher population growth compared with those at lower food levels. An interaction between toxicant and food level was evident on the population growth of E. dilatata. Results have been discussed in light of the protective role of algal density on the toxic effects of insecticides on rotifers and the differences in susceptibility to toxicants between planktonic and littoral rotifers.

Animals↗

Precopulatory mate guarding and mating behaviour in the rotifer Epiphanes senta (Monogononta: Rotifera).

Epiphanes senta is a littoral rotifer species that occurs in temporary waters and displays a mating behaviour which has not, to my knowledge, so far been described for monogonont rotifers. Monogonont rotifers show distinctive periods within their life cycle during which mictic females appear. Mictic females produce haploid eggs that develop into males or into diapausing eggs if fertilized. The females of E. senta are mostly stationary on the substrate while males are more active swimmers. If they encounter eggs with female embryos of their own species, they attend them and mate with the hatching female. Experiments showed that males are able to discriminate between male, female and diapausing eggs. They exhibit a strong preference for female eggs that are only a few hours away from hatching compared with eggs in early developmental stages. Further experiments did not show any significant differences in male attendance of mictic and amictic eggs. It is hypothesized that males judge the age of a female egg by sensing a chemical that is produced by the growing embryo and diffuses through the egg shell. The male mating behaviour is similar to precopulatory mate guarding known from arthropods but it lacks the monopolization of the female by the male.

Analysis of Variance↗

Effect of three food types on the population growth of Brachionus calyciflorus and Brachionus patulus (Rotifera: Brachionidae).

We compared the population growth of B. calyciflorus and B. patulus using the green alga Chlorella vulgaris, baker's yeast Saccharomyces cerevisiae or their mixture in equal proportions as food. Food was offered once every 24 h in two concentrations (low: 1 x 10(6) and high: 3 x 10(6) cells ml-1) separately for each species. The experiments were terminated after 15 days. In general, at any food type or concentration, B. patulus reached a higher population density. A diet of Chlorella alone supported a higher population growth of both rotifer species than yeast alone. B. calyciflorus and B. patulus achieved highest population densities (103 +/- 8 ind. ml-1 and 296 +/- 20 ind. ml-1, respectively) on a diet of Chlorella at 3 x 10(6) cells ml-1. When cultured using the mixture of Chlorella and yeast, the maximal population densities of B. calyciflorus were lower than those grown on Chlorella. Under similar conditions, the maximal abundance values of B. patulus were comparable in both food types. Regardless of food type and density the rate of population increase per day (r) for B. calyciflorus varied from 0.13 +/- 0.03 to 0.63 +/- 0.04. These values for B. patulus ranged from 0.19 +/- 0.01 to 0.37 +/- 0.01. The results indicated that even though Chlorella was a superior food for the tested rotifers, yeast can be effectively used at low concentrations to supplement algal requirements in rotifer culture systems.

Animals↗

Toxicity testing using esterase inhibition as a biomarker in three species of the genus Lecane (Rotifera).

We have developed an esterase inhibition test to investigate the effects of 10 toxicants, including six metals (cadmium, chromium, copper, lead, mercuric chloride, and titanium) and four organics (benzene, ethyl acetate, toluene, and vinyl acetate) in three species of the benthic rotifer genus Lecane (L. hamata, L. luna, and L. quadridentata). Metals affect esterase inhibition by an average value of 4,957-fold greater than the four organics tested for the three rotifer species. Most of the EC50 (effect concentration where a 50% reduction in esterase activity is observed) values correspond to environmentally realistic concentrations. Comparisons of acute-to-chronic ratios among these three species showed that in two species, L. luna and L. quadridentata, esterase inhibition is an outstanding biomarker for most of the toxicants tested.

Animals↗

Effect of different densities of live and dead Chlorella vulgaris on the population growth of rotifers Brachionus calyciflorus and Brachionus patulus (Rotifera).

In order to maintain rotifer populations during periods of low algal production, it is necessary to offer alternate diets, some of which include forms of preserved algae. The present work is based on the effect of live and dead Chlorella vulgaris on the population growth of Brachionus calyciflorus and Brachionus patulus. The experimental design consisted of three algal levels (0.5 x 10(6), 1.5 x 10(6) and 4.5 x 10(6) cells ml-1) offered in three forms (living, frozen and heat-killed). The maximal population density values for B. calyciflorus ranged from 55 +/- 1 ind. ml-1 (at 0.5 x 10(6) cells ml-1) to 471 +/- 72 ind. ml-1 (at 4.5 x 10(6) cells ml-1) with live Chlorella, but was much lower (6 +/- 1 to 26 +/- 6 ind. ml-1) with frozen or heat-killed alga under comparable food levels. However, the maximum population density of B. patulus under live or or heat-killed Chlorella was similar at comparable algal levels but when offered frozen algae it was four times less. The highest mean peak population density was 1,277 +/- 83 ind. ml-1 under 4.5 x 10(6) cells ml-1. The rate of population increase for B. calyciflorus varied from 0.50 to 0.79 using live Chlorella, but under comparable conditions, this range was lower (0.21 to 0.31) for B. patulus. Results have been discussed in light of possible application for aquaculture.

Animal Feed↗