Demographic parameters of Brachionus patulus Muller (Rotifera) exposed to sublethal DDT concentrations at low and high food levels

The combined effects of sublethal levels of DDT (0, 15, 30, 45, and 60 µg·l^–1) and food (Chlorella at 1 and 3 × 10⁶ cells ml^–1) on the demography of the rotifer Brachionus patulus were studied. The average lifespan, life expectancy, net reproductive rate (R_o) and intrinsic rate of increase (r) were significantly greater at the higher food level, but declined with increasing DDT concentration. A significant DDT food interaction was manifested in the toxicity-mitigating action of the higher food density at higher DDT levels. The r values were negative at DDT levels 30 µg · l^–1 and above, only when food density was low. The effective DDT concentration at which a given parameter value was reduced to 50% of that in the controls (EC50) was lower at lower food levels for both survivorship and reproduction, and lower for reproduction than for survivorship at either food level.     

Heterologous expression and secretion of sweet potato peroxidase isoenzyme A1 in recombinant Saccharomyces cerevisiae

A vector system has been developed to express isoenzyme A1 of sweet potato peroxidase (POD) and was introduced into Saccharomyces cerevisiae. The system contains the signal sequence of Aspergillus oryzae -amylase to facilitate the extracellular secretion of peroxidase under the control of constitutive glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter. In a batch culture using YNBDCA medium (yeast nitrogen base without amino acids 6.7 g l^–1, Casamino acids 5 g l^–1 and glucose 20 g l^–1), the recombinant strain expressed the swpa1 gene giving a secretion yield of POD activity of ca. 90% of total expressed peroxidase. Supplementation with PMSF (0.05 mM) and Casamino acids (5 g/50 ml) increased extracellular POD activity to nearly 10 kU ml^–1, equivalent to 1.5 kU g^–1 cell dry wt. This is 9 fold higher than that obtained in medium without PMSF. From SDS-PAGE and native-PAGE analyses POD has an M _r of 53 kDa.     

Effect of salinity on competition between the rotifers Brachionus rotundiformis Tschugunoff and Hexarthra jenkinae (De Beauchamp) (Rotifera)

We studied the effect of different concentrations (0, 3, 6, 9 and 12 g l^–1) of sodium chloride at one food level of Chlorella (1×10⁶ cells ml^–1) on competition between the rotifers B. rotundiformis and H. jenkinae, both of which were isolated from a saline lake. The population growth experiments were conducted for 3 weeks. Both the rotifer species did not survive beyond one week at a salinity of 0 g l^–1. Regardless of salt concentration and the presence of a competitor, H. jenkinae reached higher densities than B. rotundiformis. When grown alone, both B. rotundiformis and H. jenkinae showed optimal peak population densities at the salinity of 6 and 9 g l^–1. Since biomass wise, B. rotundiformis was larger than H. jenkinae, it showed a lower numerical abundance. Thus, the maximum peak population densities of B. rotundiformis and H. jenkinae recorded in this study were 107±3 and 203±28 ind. ml^–1. The maximal rates of population increase for B. rotundiformis and H, jenkinae when grown alone were 0.264±0.003 and 0.274±0.004, respectively. Our results also indicated that B. rotundiformis and H. jenkinae coexisted better at a salinity of 6 and 9 g l^–1 of sodium chloride while a salinity of 3 g l^–1 favoured Hexarthra over B. rotundiformis. At 12 g l^–1, both the rotifer species grown alone or together showed lower growth rates compared to those at lower salinity levels. Except 0 g l^–1, in all other salinity treatments, H. jenkinae was a superior competitor to B. rotundiformis.     

Culture conditions of tannase production by Lactobacillus plantarum

Lactobacillus plantarum produced an extracellular tannase after 24 h growth on minimal medium of amino acids containing 2 g tannic acid l^–1. Enzyme production (6 U ml^–1) was optimal at 37 °C and pH 6 with 2 g glucose l^–1 and 7 g tannic acid l^–1 in absence of O₂.     

Brachionus calyciflorus Pallas as agent for the removal of E. coli in sewage ponds

Brachionus calyciflorus (Pallas) is a common brachionid in sewage oxidation ponds. The uptake and assimilation of E. coli was optimal at concentrations of 2.7–6.9 × 10⁸ cells ml^–1 while assimilation coefficient per body weight of B. calyciflorus was found to be 10% · Ind.^–1 d^–1. More than two eggs per individual were produced during 24 hours when brachionids were fed with a mixutre of E. coli (10⁹ cells · ml^–1) and Chlorella spp. (10⁶ cells · ml^–1). The nutritional value of the mixture of E. coli and Chlorella spp. was found to be higher than that of bacteria alone.     

Life table demography and population growth of Brachionus variabilis Hempel, 1896 in relation to Chlorella vulgaris densities

We studied the life history variables and population growth characteristics of Brachionus variabilis, which was recorded for the first time from Mexico. The animals were fed Chlorella, using five concentrations (0.25, 0.5, 1, 2 and 4 × 10⁶ cells ml^–1) at 25 °C. Food density was observed to have significant effect on life expectancy, average lifespan, gross reproductive rate, net reproductive rate, generation time and population growth rate. The average lifespan ranged from 3 to 6 days depending on the food density. The net reproductive rate ranged from 2 to 7 neonates female^–1 d^–1. The rate of population increase per day varied from 0.14 to 0.35. The highest net reproductive rate and average lifespan and life expectancy were recorded at Chlorella concentrations of 1 × 10⁶ and 2 × 10⁶ cells ml^–1.     

Production and nutritional quality of the rotifer Brachionus plicatilis fed marine Chlorella sp. at different cell densities

The effect of different cell densities of marine Chlorella sp. on the growth rate, doubling time and production of the rotifer Brachionus plicatilis was investigated. A significant increase in rotifer production was achieved at a density of 50 × 10⁶ Chlorella cells ml^–1. The nutritional quality of rotifers grown at different concentrations of Chlorella is discussed.     

A laboratory study of feeding and assimilation in Euchlanis dilatata lucksiana

Feeding of Euchlanis dilatata lucksiana, a brachionid rotifer isolated from the Loosdrecht Lakes (The Netherlands), was examined in the laboratory using ¹⁴C-labelled food. The gut-filling time at a food concentration of 9.6 µg C ml^–1 was about 15 minutes. Animals which were fed on 3 size fractions of the lake seston (< 7 µm, 7–15 µm, and 15–33 µm) showed a clear preference (Jacobs' selectivity index) for the largest size fraction. This fraction was composed predominantly of filamentous cyanobacteria. For animals weighing 0.37–0.49 µm C ind.^–1 the daily ration (daily food consumption per unit body weight × 100) ranged from 50 to 100% at food levels of 2 mg C l^–1 and below, but increased to 150–250% at food concentrations of 5 mg C l^–1 and above. The assimilation efficiency was 100% up to 5 mg C l^–1 of food, but decreased to about 80% at higher food levels.     

High density culture of the freshwater rotifer,Brachionus calyciflorus

The freshwater rotifer, Brachionus calyciflorus is one of the live food organisms used for the mass production of larval fish. In this study possibility of obtaining high density cultures of the freshwater rotifer B. calyciflorus were investigated. The two culture systems used differed in their air and dissolved oxygen supplies using three temperatures in each case: 24, 28 and 32 °C. Rotifers were batch-cultured using 5 l-vessels and fed with the freshwater Chlorella. The growth rate of rotifers significantly increased with an increase in temperature. The maximum density of the rotifers with air-supply at 24 °C, 6500 ind. ml^–1, was significantly lower than those cultured at 28 and 32 °C, i.e. 8600 and 8100 ind. ml^–1, respectively. Dissolved oxygen levels decreased with time and ranged from 0.8 to 1.4 mg l^–1 when the density of freshwater rotifer was the highest at each temperature. The highest density (19200 ind. ml^–1) of freshwater rotifer was obtained in cultures with a supply of oxygen at 28 °C. Densities of 13500 and 17200 ind. ml^–1 were found at 24 and 32 °C, respectively. Levels of NH₃-N increased with time and a dramatic increase of NH₃-N was observed at high temperatures. Levels of NH₃-N at 24, 28 and 32 °C were 13.2, 18.5 and 24.5 mg l^–1, respectively. These levels coincided with the highest rotifer density at each of the three temperatures. When rotifers were cultured with an oxygen-supply and pH was adjusted to 7, the maximum density of rotifer reached 33500 ind. ml^–1 at 32 °C . These results suggested that high density culture of freshwater rotifer, B. calyciflorus could be achieved under optimal conditions with DO value of exceeding 5 mg l^–1 and NH₃-N values of lower than 12.0 mg l^–1.     

Preparation and testing of Sardinella protein hydrolysates as nitrogen source for extracellular lipase production by Rhizopus oryzae

Various fish protein hydrolysates (FPH) from sardinella (Sardinella aurita) were used as nitrogen sources for the production of extracellular lipase by the filamentous fungus Rhizopus oryzae. The best results were obtained with defatted meat–fish protein hydrolysates (DMFPH), indicating the presence in the lipid fraction of some constituents which may repress lipase synthesis. Furthermore, it was found that the extensive hydrolysis of fish proteins resulted in a higher lipase production. The use of 40 g DMFPH l^–1 for the growth of Rhizopus oryzae in medium R1 resulted in a lipase production of 394 U ml^–1, higher than the yield obtained with standard soy peptone as nitrogen source (373 U ml^–1). The most appropriate medium for the growth and the production of lipase is composed only of 24 g DMFPH l^–1 and 10 g glucose l^–1, indicating that the strain can obtain its nitrogen and salts requirements directly from fish substrate.     

Influence of culture temperature on the growth,biochemical composition and fatty acid profiles of six Antarctic microalgae

The growth, biochemical composition and fatty acid profiles of six Antarctic microalgae cultured at different temperatures, ranging from 4, 6, 9, 14, 20 to 30 ^∘C, were compared. The algae were isolated from seawater, freshwater, soil and snow samples collected during our recent expeditions to Casey, Antarctica, and are currently deposited in the University of Malaya Algae Culture Collection (UMACC). The algae chosen for the study were Chlamydomonas UMACC 229, Chlorella UMACC 234, Chlorella UMACC 237, Klebsormidium UMACC 227, Navicula UMACC 231 and Stichococcus UMACC 238. All the isolates could grow at temperatures up to 20 ^∘C; three isolates, namely Navicula UMACC 231 and the two Chlorella isolates (UMACC 234 and UMACC 237) grew even at 30 ^∘C. Both Chlorella UMACC 234 and Stichococcus UMACC 238 had broad optimal temperatures for growth, ranging from 6 to 20 ^∘C (μ = 0.19 – 0.22 day^–1) and 4 to 14 ^∘C (μ = 0.13 – 0.16 day^–1), respectively. In contrast, optimal growth temperatures for NaviculaUMACC 231 and Chlamydomonas UMACC 229 were 4 ^∘C (μ = 0.34 day^–1) and 6–9 ^∘C (μ = 0.39 – 0.40 day^–1), respectively. The protein content of the Antarctic algae was markedly affected by culture temperature. All except Navicula UMACC 231 and Stichococcus UMACC 238 contained higher amount of proteins when grown at low temperatures (6–9 ^∘C). The percentage of PUFA, especially 20:5 in Navicula UMACC 231 decreased with increasing culture temperature. However, the percentages of unsaturated fatty acids did not show consistent trend with culture temperature for the other algae studied.     

Tannase production by Bacillus licheniformis

Bacillus licheniformis KBR 6 produced maximum extracellular tannase activity at 0.21 U ml^–1 with 1.5% (w/v) tannic acid either in the absence or presence of glucose (1 g l^–1) after 18–21 h growth though the organism did not attain maximum growth until 36 h.     

Improvement of nisin production in pH feed-back controlled,fed-batch culture by <Emphasis Type="Italic">Lactococcus lactis</Emphasis>subsp<Emphasis Type="Italic">. lactis</Emphasis>

Nisin production by Lactococcus lactis subsp. lactisin fed-batch culture was doubled by using a pH feed-back controlled method. Sucrose concentration was controlled at 10 g l^–1 giving 5010 IU nisin ml^–1 compared to 2660 IU nisin ml^–1 in batch culture.     

Production of cellulases in batch culture using a mutant strain of Trichoderma reesei growing on soluble carbon source

Trichoderma reesei Rut-C30 is a highly derepressed mutant which synthesised active cellulases in culture media containing glucose and lactose as the only carbon sources. The maximum biomass, filter paper and specific filter paper activities for cell growth on 20 g glucose l^–1 were 20 g dry cell wt l^–1, 1.9 FPU ml^–1 and 4.8 FPU mg^–1 protein respectively, while on 40 g glucose l^–1 were 25 g dry cell wt l^–1, 4.5 FPU ml^–1 and 6.2 FPU mg^–1 protein, respectively. This strain had a higher specific filter paper activity (6.2 FPU mg^–1 protein) than was produced by other T. reesei mutants (3.6 FPU mg^–1 protein).     

The efficacy of Scenedesmus morphology as a defense mechanism against grazing by selected species of rotifers and cladocerans

Phytoplankton often develop various defense mechanisms in response to zooplankton grazing, such as spines and colonies. While it is now known that increased spine length and cells in a colony of members of the genus Scenedesmus, when zooplankton grazing is intense, helps in reducing zooplankton filtering rates, the effect of these defense mechanisms at the population level has been observed in few studies. Here we present data on the growth rates of four zooplankton species, Brachionus calyciflorus, B. patulus, Ceriodaphnia dubia and Daphnia pulex at two food levels using two species of colony-forming Scenedesmus spp.: S. acutus (cell length = 18.2 ± 0.4 µm; width = 4.2 ± 0.1 µm; average colony length = 90 µm; width: 21 µm) and S. quadricauda (cell length: 21 ± 0.5 width 7.5 ± 0.3 µm; average colony length: 84 µm; width: 30 µm). Whereas S. acutus had no spines, S. quadricauda had spines of 6–10 µm. Population growth experiments of the test rotifers and cladocerans were conducted in 100 ml containers with 50 ml of the medium with test algae. Algae concentrations used were: 13 and 52 mg dw l^–1 of each of the two algal species offered in colonial forms. We used an initial inoculation zooplankter density of 1 ind. ml^–1 for either of the rotifer species and 0.2 ind. ml^–1 for either of the cladoceran species. In all, we had 64 test containers (4 test species of zooplankton × 2 test species of algae × 2 algal densities × 4 replicates). We found a significant effect of algal size on the growth rates of all the four tested species of zooplankton. The population growth rates of zooplankton ranged from –0.58 to 0.66 and were significantly higher on diet of S. acutus than of S. quadricauda. Thus, our study confirms that the larger colony size and the formation of spines in S. quadricauda were effective defenses against grazing by both rotifers and smaller sized cladoceran Ceriodaphnia dubia but that larger-bodied Daphnia pulex could exploit both the algal populations equally.     

Seasonal succession of the phytoplankton in the upper Mississippi River

Species composition and seasonal succession of the phytoplankton were investigated on the upper Mississippi River at Prairie Island, Minnesota, U.S.A. Both the numbers and volume of individual species were enumerated based on cell counts with an inverted microscope. A succession similar to algal succession in the local lakes occurred. The diatoms were dominant during the spring and fall and blue-green algae were dominant during the summer. The algal concentrations have increased up to 40 fold the concentrations of the 1920's, since the installation of locks and dams. The maximum freshweight standing crop was 4 mg · l^–1 in 1928 (Reinhard 1931), 13 mg · l^–1 in 1975 a wet year, and 47 mg · l^–1 in 1976, a relatively dry year with minimal current discharge. The diatoms varied from 36–99%, the blue-green algae from 0–44% and the cryptómonads from 0–50% of the total standing crop. The green algae were always present but never above 21% of the biomass. The dominant diatoms in recent years were centric -Stephanodiscus andCyclotella spp. (maximum 50,000 ml^–1). The dominant blue-green algae wereAphanizomenon flos-aquae (L.) Ralfsex Born.et Flahault andOscillatoria agardhii Gomont (maximum 800 ml^–1). These algal species are also present in local lakes. Shannon diversity values indicated greatest diversity of algae during the summer months.     

Production of a thermostable extracellular lipase by Kluyveromyces marxianus

Kluyveromyces marxianus was grown in submerged culture in a complex medium with several potential inducers of lipolytic activity (triacylglycerols, fatty acids). The highest extracellular lipolytic enzyme production (about 80 U ml^–1 in 3 d) was obtained when the medium was supplemented with 2 g urea l^–1 plus 5 g tributyrin l^–1. Addition of surfactants (1 g l^–1) did not improve production. The lipase had a high thermal stability in aqueous solution (73% residual activity after 9 d at 50 °C, 16 min half-life time at 100 °C). It was also stable at acidic pH and showed good tolerance to organic solvents (70% residual activity after 2 d in n-hexane of cyclohexane).     

Effect of two microalgae concentrations on body size and egg size of the rotifer <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis>

The rotifer, Brachionus calyciflorus, was grown with two algae species (Chlorella sp. and Scenedesmus obliquus) at different concentrations (0.1, 1 and 10 × 10⁶ cells ml⁻¹). The body size (lorica biovolume) of individual rotifer and their egg size were measured when the populations were roughly in the exponential phase of population growth. The body size of the rotifers differed significantly (P < 0.05) among the two algae species used, however this effect was not observed for egg size. The body size of rotifers fed on higher densities of Chlorella sp. (10 × 10⁶ cells ml⁻¹) was significantly larger than for those fed on lower and medium densities (0.1 and 1 × 10⁶ cells ml⁻¹). Body size and egg size of rotifers fed with different amounts of Scenedesmus did not differ significantly. The egg size was significantly larger at higher food level of Chlorella. A significantly positive correlation was observed between the adult rotifer body size and their egg size.     

Selective ingestion of phytoplankton by the bivalvesMytilus edulis L. andCerastoderma edule (L.)

Blue mussles (Mytilus edulis) and cockles (Cerastoderma edule) were collected in the Oosterschelde estuary (SW Netherlands) and exposed to diets consisting of the diatomPhaeodactylum tricornutum and silt. Algal concentrations were kept constant (20.10³ cells.ml^–1), and silt concentrations were varied between 3–80 mg.l^–1 (mussel experiments) and 20–120 mg.l^–1 (cockle experiments). Chlorophyll-a concentrations in the pseudofaeces were reduced compared to the diets, indicating selective ingestion of the algae. It was estimated that the mussels increase the ingestion of chlorophyll-a 2.0 times, and the cockles increase the chlorophyll-a ingestion 2.8 times, compared to the ingestion rate without selection. The selection coefficients were not affected by the SPM concentrations used. Phaeophytin-a concentrations in the faeces showed an increase as a consequence of the digestion of the ingested algae. Digestive efficiencies of chlorophyll-a varied between 36–92% The digestive efficiencies decreased with increasing SPM concentrations.Communication no. 541 of the Delta Institute for Hydrobiological Research.     

Efficient production by Escherichia coli of recombinant protein using salting-out effect protecting against proteolytic degradation

To produce glucoamylase efficiently as a recombinant protein, E. coli was grown with 20 g (NH₄)₂SO₄ l^–1 which removed proteolytic activity but did not effect cell growth. Growth in M9 medium with 20 g (NH₄)₂SO₄ l^–1 produced 11 U glucoamylase ml^–1 compared to 7 U ml^–1 without addition. Furthermore, the glucoamylase activity was maintained at about 9 U ml^–1.