Rotifers of the Kalahari Gemsbok National Park,South Africa

The Kalahari Gemsbok National Park in the Northern Cape Province, South Africa, is bounded by two dry river beds in which 39 artificial water sources have been provided for the use of the wild animals. The water in these sources varies from fresh to highly saline and an opportunity is thus provided to determine, not only which rotifer species are able to colonise such sources, but what their salinity tolerances are. The rotifer fauna, studied in summer and winter, is dominated by mixed populations of Brachionus calyciflorus, B. plicatilis and Hexarthra jenkinae, with only a few other monogononts and bdelloids. The most striking feature of the rotifer fauna in these troughs is the very low species diversity encountered.     

Presence and development of populations of the introduced brown alga Sargassum muticum in the southwest Netherlands

Of the three Brachionus species used in aquaculture, Brachionus rubens, B. calycilorus and B. plicatilis, the latter is most widely used in raising marine fish and shrimp larvae due to its tolerance to the marine environment. In freshwater aquaculture the use of B. rubens and B. calycilorus is limited, probably because inert food products are readily available as feed for freshwater larvae.The rotifer Brachionus plicatilis is used in large numbers as the first food organism in intensive cultures of marine fish and shrimp larvae. An adequate supply of these rotifers relies on mass cultures. The reproductive rate of rotifers in these cultures depends on food quality and quantity, salinity, temperature and pH of the medium. Removal of waste products from culture tanks leads to higher and more efficient production of rotifers over extended periods of time. Rotifers have to be enriched with polyunsaturated fatty acids, which are essential for proper development and survival of marine fish and shrimp larvae.The future use of preserved rotifers and their resting eggs may help to overcome unforeseen failures of live cultures and may lead to more efficient use of these organisms in raising freshwater and marine fish and shrimp larvae.     

Brachionus plicatilis tolerance to low oxygen concentrations

Tolerance to low oxygen concentrations is expected in Brachionus plicatilis, a rotifer adapted to live in saline warm waters. The population dynamics of a clone of this species, isolated from an endorreic saline lake, was studied under controlled laboratory conditions. Although their growth and metabolism is extremely reduced, B. plicatilis populations are able to maintain relatively high-density populations (a mean of 35 ind ml^–1) in oxygen concentrations below 1 mg 1^–1, for more than one month. Major features of population growth related to oxygen are discussed.     

Evaluating decline parameters of rotifer Brachionus plicatilis populations as an interstitial water toxicity bioassay

Population decline of the rotifer Brachionus plicatilis has been evaluated as a sensitive and reliable bioassay for assessing toxicity of marine sediment interstitial water. Three environmental conditions were examined using both interstitial and seawater cultures: (a) adverse effects from dissolved contaminant concentrations; (b) effects from particulate organic matter; and (c) increase or decrease of bacterial biomass from addition of mixtures of antibiotics. Three different parameters were measured to characterize decline: (i) time required for 50% of inoculated population to die (TL₅₀); (ii) curve of the decline rate of Brachionus plicatilis (µ _BP and (iii) graphical area contained below plot of the egg:female ratio curve against time (A _BPM ).Results indicated that, for each of the different assays, the three parameters of the Brachionus plicatilis population decline test were sensitive to concentrations of contaminants dissolved in interstitial waters (principally: total ammonia, Cu, Cr and alkylbenzenesulphonates) but not to particulate organic matter. Nevertheless, the presence or absence of mixed antibiotics with the contaminants may influence the Brachionus plicatilis population decline test, principally by retarding the hatching of eggs. Based on these results, Brachionus plicatilis is confirmed as an appropriate organism for use as an indicator of interstitial water contamination, using either decline rate, TL₅₀ or both parameters. The presence of particulate matter has no effect on these parameters, but the bacterial population may be an influence, although to a lesser extent than the toxicants.     

Axenic culture of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> using antibiotics

The rotifer Brachionus plicatilis culture is composed of complex microcosms including bacteria, protozoans, algae, and fungi. Previous studies reported methods to establish axenic rotifer cultures, but further refinement of these techniques is needed, for molecular biological research which requires pure culture to isolate nucleic acids from rotifers only. In order to render rotifer culture axenic, we tested five antibiotics: ampicillin (Amp), chloramphenicol (Cp), kanamycin (Km), nalidixic acid (Na), and streptomycin (Sm) at 30–100 μg/ml. Except for Cp, which reduces rotifer reproduction, all other antibiotics at the tested concentrations did not affect rotifer reproduction or show any toxic effects. A rotifer disinfection method was finally established by treating the resting eggs with 0.25% (w/v) sodium hypochlorite (NaOCl) for 3 min, washing with sterilized sea water, and then exposing the neonates to an Amp, Km, Na, and Sm mixture. Using four nutrient media, we confirmed that this protocol renders the rotifer culture bacterial and fungus free. The axenic rotifer culture generated here is useful not only for genetic analysis of Brachionus plicatilis, but for studying the rotifer life cycle without bacterial influence.     

Molecular Characterization of Mn-superoxide Dismutase and Gene Expression Studies in Dietary Restricted <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Rotifers

Superoxide dismutases (SODs) promote a conversion of harmful reactive oxygen species (ROS) to relatively moderate forms, resulting in the extension of lifespan in the nematode Caenorhabditis elegans under caloric restriction. The lifespan of the rotifer Brachionus plicatilis is also markedly extended by caloric restriction. We, therefore, cloned cDNA encoding SOD activated with Mn (Mn SOD) from B. plicatilis and examined its expression pattern in rotifers raised with energy restricted diet. The full length deduced amino acid sequence of the rotifer Mn SOD showed 61% identity with the C. elegans ortholog. Four amino acid residues that are essential to the binding of this enzyme to Mn were conserved in the rotifer Mn SOD. Subsequently we examined the mRNA expression patterns of Mn SOD using highly sensitive quantitative real-time PCR for various rotifer populations that are likely to differ in their lifespans in experiments on calorie restricted diets. The accumulated mRNA levels of Mn SOD were found to increase in supposedly long-lived rotifers. These results suggest that Mn SOD is possibly related to the aging of B. plicatilis.     

Inland salt waters of southern Africa

Inland salt lakes are widely distributed in southern Africa: they are particularly common in South Africa, but many occur in Namibia and Botswana. All are shallow, and most are ephemeral with salinities that are not very high (mostly < 50 g l⁻¹). Fringing zones of halophytes or submerged macrophytes are neither well-developed nor taxonomically diverse. The Cyanobacteria, especially Nodularia spumigena, often dominate the phytoplankton. The fauna of the Makgadikgadi area (northeast Botswana) is diverse and is similar to that of East African salt lakes. The aquatic fauna of salt water south of the Makgadikgadi Basin, on the other hand, is extremely depauperate, has no well-defined assemblage confined to saline waters, and appears mostly to comprise tolerant freshwater forms. Lovenula falcifera and Metadiaptomus transvaalensis (diaptomid copepods), Moina micrura (Cladocera) and Brachionus plicatilis (Rotifera) are frequently encountered zooplankton species, a few species of insects (Anisops sp., beetles, chironomids and ephydrids) are the principal non-planktonic macroinvertebrates. Artemia ‘salina’ is occasionally present, but may be an introduced form. The avifauna, in contrast to the aquatic macroinvertebrate fauna, is rich, with the greater and lesser flamingo often common.     

Behavioural Reproductive Isolation in a Rotifer Hybrid Zone

A hybrid zone between two Brachionus plicatilis rotifer mitochondrial DNA (mtDNA) lineages was recently described in the Iberian Peninsula between a pond (Santed 2) and a lake (Gallocanta). The patterns of mitochondrial and nuclear genetic variation observed suggested that gene flow is mainly male-mediated from the lake to the pond. Here we test two hypotheses: (a) that male-mediated gene flow occurs through assortative mating between individuals from these ponds, (b) that behavioural isolation occurs between the two mtDNA lineages. We isolated, reared and genotyped rotifer clones from resting eggs collected in the sediments of these and two other distant ponds. We devised a quick, inexpensive RFLP method to discriminate between B. plicatilis and its sibling species B. ‘Manjavaeas’ and between both mtDNA B. plicatilis lineages. Behavioural no-choice tests using new-born, virgin males and females were performed between five clones. B. ‘Manjavacas’ and B. plicatilis were reproductively isolated. B. plicatilis clones did not show evidence of reproductive isolation, regardless of their mtDNA lineage, except Santed 2 males, which discriminated strongly against Gallocanta females. These results could help to explain the discrepancies between mitochondrial and nuclear genetic variation reported in the two populations.     

Associations of planktonic and periphytic rotifers in a Malaysian estuary and two nearby ponds

Samples were taken from a Malaysian estuary along a salinity gradient from full strength sea water down to a conductivity of 27 µS cm^–1. In a region where the conductivity fluctuated between 6500 and 14000 µS only empty loricas of Keratella cochlearis and Lecane bulla were found. Where the salinity reached a maximum of 3800 µS Lecane bulla was the dominant rotifer, and 17 other species were found. Where the conductivity remained below 1000 µS at all stages of the tidal cycle there were more species of rotifers, up to 33 at one station.The rotifer fauna of the estuary was compared to the assemblage found in two local ponds. Of the 63 species found in the estuary only 29 were also found in the ponds. Compared to the total known rotifer fauna of Malaysia the estuary has a higher proportion of Lecane species (40%), and the genera Brachionus and Trichocerca are under-represented.Species associated with inland saline waters, such as Brachionus plicatilis, B. dimidiatus and Hexarthra jenkinae were not found in the estuary.     

The essential omega-3 fatty acid EPA affects expression of genes involved in the metabolism of omega-6-derived eicosanoids in Daphnia magna

We quantified the predation of Acanthocyclops americanus from the shallow Mediterranean lake Albufera, using gut contents from field collections and laboratory feeding tests. For functional response studies, we used Brachionus plicatilis (at 6 concentrations, 400–4000 ind. 40 ml⁻¹) and Diaphanosoma mongolianum (at 2–20 ind. 40 ml⁻¹). Copepod feeding rates were also estimated using different proportions of rotifer prey and lake seston (0–67.5% of seston + 40 individuals of B. plicatilis). Prey selection studies were conducted using five zooplankton species: Brachionus angularis, Brachionus plicatilis, Keratella tropica, Daphnia magna and Diaphanosoma mongolianum. Gut contents of field-collected adult Acanthocyclops contained filamentous algae and cyanobacteria and 16 zooplankton species (Keratella cochlearis, unspined and spined forms, K. tropica, Brachionus plicatilis, Brachionus calyciflorus, Brachionus angularis, Brachionus variabilis, Asplanchna girodi, Polyarthra vulgaris, Synchaeta pectinata, Lepadella rhomboides, unidentified bdelloids, Alona rectangula, Chydorus sphaericus, Bosmina longirostris, D. magna, Ceriodaphnia dubia and copepod nauplii). When fed B. plicatilis or D. mongolianum, female A. americanus had higher prey consumption rates than males. Increased proportion of lake seston caused reduced consumption of brachionid prey. Our data suggest that A. americanus is omnivorous in nature.

     

Additions to the rotifer fauna of south-east Asia: new and rare species from north-east Thailand

Ninety-three freshwater habitats in the north-eastern part of Thailand were surveyed for rotifers during March–October 1993. Two hundred species were identified, 120 of which are new to Thailand. Brachionus africanuss Segers, B. lyratus Shephard and Trichocerca hollaerti De Smet are new to Asia. Brachionus niwati n.sp. is described and figured. Brachionus donneri Brehm, Keratella edmondsoni (Ahlstrom) and Lecane blachei Berzi, endemic to south and southeast Asia, were found also.Comments are presented on some insufficiently known taxa. Of special zoogeographic interest is the record of several species previously considered endemic to Australia. An attempt is made to characterise the Thai rotifer fauna in the light of recent developments in rotifer zoogeography.     

The species composition of rotifers in the water reservoirs of Central Vietnam

The rotifer fauna has been studied in four water reservoirs in the Khanh Hoa Province, Central Vietnam. A total of 65 species and subspecies and 22 genera representing 15 families have been found; 25 species and subspecies are identified as new to Vietnam. Morphometric characteristics are given for Filinia camascela Myers, Brachionus donneri Brehm, and B. dichotomus reductus Koste et Shiel. The rotifer fauna of Central Vietnam and those of other countries of Southeast Asia are compared. Some of the tropical and subtropical species recorded have been repeatedly reported from Russia.     

Local adaptation in populations of a Brachionus group plicatilis cryptic species inhabiting three deep crater lakes in Central Mexico

1. Salinity is a strong selective force for many aquatic organisms, affecting both ecological and evolutionary processes. Most of our knowledge on the effects of salinity on rotifers in the Brachionus plicatilis species complex is based mainly on populations from waterbodies that experience broad environmental changes both seasonally and annually. We tested the hypothesis that, despite the supposedly high potential for gene flow among rotifers inhabiting neighbouring environments, constant salinity has promoted local adaptation, genetic population divergence and even cryptic speciation in B. plicatilis complex populations from three deep maar lakes of distinct salinities [1.1, 6.5 and 9.0 g L^?1 total dissolved solids (TDS)] in Central Mexico. 2. To look for local adaptation, we performed common garden experiments to test the effect of different salinities on population density and intrinsic growth rate (r). Then, we evaluated the genetic divergence by sequencing the cytochrome c oxidase subunit I (COI) gene and performed reproductive trials to assess the potential gene flow among the three populations and with other closely related B. plicatilis complex species. 3. We confirmed that the rotifer populations have phenotypic plasticity in tolerance of salinity, but only rotifers from the least saline lake are adapted to low salinity. Among the populations, sequence divergence at COI was very low (just a single haplotype was found), suggesting a persistent founder effect from a relatively recent single colonisation event and a subsequent dispersal from one lake to the others, and a very restricted immigration rate. In the phylogenetic analysis, rotifers from this area of Mexico clustered in the same clade with the middle‐sized species Brachionus ibericus and B. sp. ‘Almenara’. Mexican rotifers showed successful recognition, copulation and formation of hybrids among them, but interpopulation breeding with the Spanish B. ibericus and B. sp. ‘Almenara’ was unsuccessful. 4. We conclude that the B. plicatilis complex populations from these three lakes belong to a new biological species not yet described (presently named B. sp. ‘Mexico’). To our knowledge, this is the first report of local adaptation of a natural B. plicatilis complex population living in freshwater conditions (1.1 g L^?1 TDS).     

Nutritional effect of freshwater Chlorella on growth of the rotifer Brachionus plicatilis

Mass production of Brachionus plicatilis is usually accomplished by feeding so-called marine Chlorella (Nannochloropsis oculata) to the rotifers in marine fish hatcheries. If the marine Chlorella are in short supply, baker's yeast is usually used as a supplementary food. Recently, a condensed suspension of freshwater Chlorella (Chlorella vulgaris, k-22) was commercially developed as another supplementary food. We have evaluated the dietary value of this freshwater Chlorella for growth of the rotifer by means of individual and batch cultures. Rotifers cultured with the freshwater Chlorella suspension under almost bacteria-free conditions, showed very suppressed growth. However if the Chlorella was supplemented with vitamin B₁₂ by adding the vitamin solution into the suspension or by culturing the Chlorella in a medium containing vitamin B₁₂, the nutritional value of freshwater Chlorella was greatly improved and almost at the same level as that of marine Chlorella. Condensed Chlorella may therefore be effective as a supplementary food if vitamin B₁₂ is supplied.     

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.     

Toxicity and haemolytic activity of a newly described dinoflagellate,Heterocapsa bohainensis to the rotifer Brachionus plicatilis

The algae Heterocapsa bohaiensis is a newly described species of dinoflagellate associated with Penaeus japonicus and larvae of Eriocheir sinensis in a coastal pond of Liaodong Bay China. The rotifer Brachionus plicatilis is used as live feed for aquaculture organisms including prawns and crabs larvae. To evaluate the potential toxicity of H. bohaiensis, the effects on Brachionus plicatilis and haemolytic activity were investigated in this study. The results showed that H. bohaiensis had significant toxic effect on B. plicatilis, and this effect was dependent on the cell concentration. Significant rotifer growth suppression was observed in the ruptured cells of H. bohaiensis with ultrasonic. Relatively similar rotifer mortalities were induced both in the light and in the dark. Interestingly, haemolysis to erythrocytes was also caused in a cell density-dependent and time-dependent manner, which meant the results of haemolytic activity were consistent with the toxicity. Therefore, haemolytic toxins were considered to be involved in the toxic mechanism of H. bohaiensis against rotifers. Then, the concentrations of calcium were measured in the mastax, stomach and ovary of B. plicatilis. Obviously increased fluorescence intensity was found in the stomach, which indicated the alteration of calcium homeostasis and membrane permeability after ingesting H. bohaiensis. These results implicated haemolytic activity as a causative factor linked to the toxicity of H. bohaiensis against B. plicatilis. The results contributed to research the production and control of H. bohaiensis toxins.     

Effect of Food Quantity and Quality on Population Growth Rate and Digestive Activity in the Euryhaline Rotifer Brachionus plicatilis Müller

We investigated the nutritional effects of both food quantity and quality on Brachionus plicatilis. Decomposition of particulate and dissolved organic matter by rotifer digestive enzymes play a crucial role in rotifer nutrition. Among other enzymes, rotifers produce phosphatases, non‐specific enzymes that allow for the release of orthophosphate from a variety of organic phosphorus compounds. Phosphatase saturation was measured in B. plicatilis homogenates using the spectrofluorimetric method. We examined population growth rate, reproduction and phosphatase activity in the homogenate of rotifers (PA_RH) fed by nutrient‐replete algal food supplied at different quantities. Population growth rate, number of eggs per individual and PA_RH were affected by food quantity. Growth rate and number of eggs per individual significantly increased in rotifers fed by food supplied at the highest quantity. The highest population growth rate was reached by rotifers fed by nutrient‐replete food, while it did not significantly differ between rotifers fed on nitrogen (N)‐depleted and phosphorus (P)‐depleted food. The number of eggs per individual was more affected by N than P supply. PA_RH and rotifer RNA content were not influenced by different food quality. The results indicate that B. plicatilis is not able to regulate its digestive apparatus in terms of efficiently getting access to essential nutrients when scarce, but do this when nutrient‐replete food is available in different quantity. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Interactions between red tide microalgae and herbivorous zooplankton: effects of two bloom-forming species on the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> (O.F. Muller)

Experiments were carried out to investigate interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal bloom (HAB) species using single and mixed culture methods. B. plicatilis populations and the growth of two algae were compared at different algal cell densities. The results demonstrate that B. plicatilis obtained sufficient nutrition from Alexandrium tamarense to support net population increase. When exposed to a density of 8 × 10⁴ cells ml⁻¹ A. tamarense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (16 × 10⁴, 24 × 10⁴, 32 × 10⁴, and 40 × 10⁴ cells ml⁻¹). Cell densities of A. tamarense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on the B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, the B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of a control without addition of rotifers. Mixed culture experiments showed that A. tamarense could partly counteract the effect of H. akashiwo in limiting the rate of population increase of rotifer. In addition, the effect of different initial cell densities on interspecific competition between A. tamarense and H. akashiwo in mixed culture(s) was also investigated. The results show that A. tamarense competed very successfully when the inoculation proportions of A. tamarense and H. akashiwo were 40:5 and 40:30. Handling editor: D. Hamilton     

Population growth dynamics of the rotifer Brachionus plicatilis cultured in non-limiting food condition

Population growth parameters in batch culture of Brachionus plicatilis under a continuous supply of freeze-dried microalgae powder have been determined. Two B. plicatilis strains (L- and S-types) and four microalgae species (Nannochloropsis oculata, Nannochloropsis gaditana, Nannochloris oculata and Tetraselmis suecica) have been tested, establishing the dynamics of growth at different daily food rations. Cultures showed a short lag phase, an exponential growth phase, a long post-exponential growth phase and long decline with episodic increases. In both rotifer strains, the best growth was obtained with Nannochloropsis oculata and the poorest with Nannochloris oculata.