Palynological analysis of five selected <Emphasis Type="Italic">Onosma</Emphasis> taxa

Pollen size and exine structure of the following five Onosma L. species from Slovakia and Italy were studied by light and scanning electron microscopy: Onosma echioides, O. tornensis, O. visianii, O. arenaria, and O. pseudoarenaria. Among the taxa studied, O. echioides has the smallest pollen grains in average (10.5–17.5 × 8.0–14.5 μm), and O. pseudoarenaria the largest grains in average (15.3–20.5 × 12.3–16.3 μm). Pollen grains of all 5 species are small sized, 3-syncolporate, subprolate, heteropolar, with ovate equatorial outlines and circular to rounded triangular polar outlines; the tectum is microechinate. A positive correlation is found between pollen size and chromosome number. The value of pollen characters for taxonomic purposes and the position of the taxa studied within the genus Onosma are discussed.     

Ambrosia pollen in the air of Lublin, Poland

Studies on Ambrosia pollen concentrations were carried out in Lublin in the period 1995–2004. The effects of a number of meteorological factors were analysed. In the first period of the study, the gravimetric method was used (1995–1999), while in the second period, the volumetric method was applied. The results show an increasing trend in the amount of airborne pollen. The Ambrosia pollen season in Lublin lasts from August to October. Over a period of 5 years, the highest number of pollen grains was recorded in September (53%), followed by August (44%) and October (3%). There were wide variations in annual totals. The annual total pollen counts was 167–1180 grains, with the peak value in 2002. Maximum daily pollen concentrations (56–312 pollen grains m⁻³) were recorded in the first half of August and in the first half of September. On the days when high Ambrosia pollen concentrations occurred, the temperature was above 21°C and the winds were mainly from the southeast, south and east. Maximum intradiurnal concentrations of pollen grains occurred in the afternoon hours. These results indicate, to some degree, that Ambrosia pollen is transported for long distances before descent.     

Relationship between indoor and outdoor airborne fungal spores, pollen, and (1→3)-β-D-glucan in homes without visible mold growth

In this exploratory study, indoor and outdoor airborne fungal spores, pollen, and (1→3)-β-D-glucan levels were determined through long-term sampling (24-h) using a Button Personal Inhalable Aerosol Sampler. The air samples were collected in five Cincinnati area homes that had no visible mold growth. The total count of fungal spores and pollen in the collected samples was conducted under the microscope and Limulus Amebocyte Lysate (LAL) chromogenic assay method was utilized for the determination of the (1→3)-β-D-glucan concentration. For the combined number concentration of fungal spores and pollen, the indoor and outdoor geometric mean values were 573 and 6,435 m⁻³, respectively, with a geometric mean of the Indoor/Outdoor (I/O) ratio of .09. The geometric means of indoor and outdoor (1→3)-β-D-glucan concentrations were .92 and 6.44 ng m⁻³, respectively, with a geometric mean of the I/O ratio equal to .14. The I/O ratio of (1→3)-β-D-glucan concentration was found to be marginally greater than that calculated based on the combined number concentration of fungal spores and pollen. This suggests that (1→3)-β-D-glucan data are affected not only by intact spores and pollen grains but also by the airborne fragments of fungi, pollen, and plant material, which are ignored by traditional enumeration methodologies. Since the (1→3)-β-D-glucan level may elucidate the total exposure to fungal spores, pollen, and fungal fragments, its I/O ratio may be used as a risk marker for mold and pollen exposure in indoor environments.     

Pollen-mediated gene flow in Himalayan long needle pine (<Emphasis Type="Italic">Pinus roxburghii</Emphasis> Sargent)

Pollen migration in Pinus roxburghii has been estimated under two different conditions, viz. isolated single trees and at the fringe of natural forest. Trapping of pollen grains was conducted at geometrically increasing distances, i.e., 0, 5, 10, 20, 40, 80, 160, 320, 640, 1280 and 2560 m from the source. The observations revealed that the scattering of pollen grains was not uniform in all directions and that they travelled up to 2.5 km towards the downhill direction, while in the uphill direction dispersion of pollen grains was restricted to 320 m from the source. Pollen frequency declined rapidly as the distance from the source increased, with the highest densities within 50–100 m.     

An observation study of airborne pollen fall in Didim (SW Turkey): years 2004–2005

Pollen grains in the atmosphere of Didim, collected using a Durham sampler, were investigated in 2004 and 2005. Weekly pollen grains per square centimetre were calculated. Over a period of 2 years, 17,518 pollen grains/cm² belonging to 40 taxa and unidentified pollen grains were recorded. In 2004, 9,879 pollen grains were counted per cm², and in 2005 the value was 7,639 per cm². The majority of pollen grains investigated were Pinus spp. (45.58%), Cupressaceae/Taxaceae (13.49%), Olea spp. (9.19%), Platanus spp. (7.62%), Gramineae (6.33%), Pistacia spp. (4.34%), Morus spp. (3.81%), Quercus spp. (2.02%), Abies spp. (1.39%), and Plantago spp. (1.11%). During the month of April, 40.46% of total pollen grains were recorded. According to our results, pollen season durations for the dominated pollen grains in Didim were: the 7th–33rd weeks for Pinus spp., nearly the whole year except summer for Cupressaceae/Taxaceae, the 17th–29th weeks for Olea spp., the 10th–24th weeks for Platanus spp., the 8th–46th weeks for Gramineae, the 8th–20th weeks for Pistacia spp., the 11th–21st weeks for Morus spp., the 17th–21st weeks for Quercus spp., the 9th–27th weeks for Abies spp., and the 7th–26th weeks for Plantago spp.     

The pollen season dynamics and the relationship among some season parameters (start,end, annual total,season phases) in Kraków,Poland, 1991–2008

The dynamics of 15 taxa pollen seasons in Kraków, in 1991–2008 was monitored using a Burkard volumetric spore trap of the Hirst design. The highest daily pollen concentrations were achieved in the first half of May, and they were caused mainly by Betula and Pinus pollen. The second period of the high concentrations took place from the middle of July to the end of August (mainly Urtica pollen). Tree pollen seasons were shorter (18–24 days) in comparison with the most herbaceous pollen seasons (73–89 days), except at Artemisia and Ambrosia seasons (30 and 24 days, respectively). The season phases (percentyles) of the spring and late-summer taxa were the most variable in the consecutive years. The highest annual sums were noted for Urtica, Poaceae (herbaceous pollen seasons) and for Betula, Pinus, Alnus (tree pollen seasons), and the highest variability of annual totals was stated for Urtica, Populus, Fraxinus and the lowest for Ambrosia, Corylus, Poaceae. For the plants that pollinate in the middle of the pollen season (Quercus, Pinus and Rumex), the date of the season start seems not to be related to the season end, while for late pollen seasons, especially for Ambrosia and Artemisia, the statistically negative correlation between the start and the end season dates was found. Additionally, for the most studied taxa, the increase in annual pollen totals was observed. The presented results could be useful for the allergological practice and general botanical knowledge.     

Pinus pollen in the atmosphere of Vigo and its relationship to meteorological factors

The Pinus genus has an elevated pollen production and an anemophilous nature. Although considered to be hypoallergenic, numerous cases of allergies caused by Pinus pollen have been cited and different authors believe that its allergenicity should be studied in more depth. In the city of Vigo several patients have tested positive for Pinus pollen extracts in skin tests, some of them being mono-sensitive to such pollens. In order to ascertain the behaviour of Pinus pollen and its correlation to the main meteorological factors, we carried out an aerobiological study in the city of Vigo from 1995 to 1998 by using a Hirst active-impact volumetric sporetrap, model Lanzoni VPPS 2000, placed on the left bank of the Vigo estuary (42°14’15’’N, 8°43’30’’W). Pinus has high quantitative importance in the airborne pollen spectrum of the city. It is one of the best represented taxa constituting 13%–20% of the total annual pollen levels. The quantity of Pinus pollen present in the atmosphere of the city of Vigo throughout a year is 5751 grains (as the average for the sampled years), with a very long pollination period, from the middle of January until May. The maximum concentration was recorded in 1998 with 1105 grains/m³ on 3 March, a much greater value than those for the previous years. At the end of its pollination period there is usually a final increase in Pinus pollen concentrations coinciding with the pollination of Pinus silvestris, which are more abundant in mountainous areas far from the city. Received: 17 March 1999 / Revised: 20 December 1999 / Accepted: 20 December 1999     

Airborne Pinus pollen in the atmosphere of Brisbane, Australia and relationships with meteorological parameters

Relationships between weather parameters andairborne pollen loads of Pinus inBrisbane, Australia have been investigated overthe five-year period, June 1994–May 1999.Pinus pollen accounts for 4.5% of the annualairborne pollen load in Brisbane where thePinus season is confined to the winter months,July–early September. During the samplingperiod loads of 11–>100 grains m³ wererecorded on 24 days and 1–10 grains m³ on204 days. The onset and peak dates wereconsistent across each season, whereas the enddates varied. The onset of the Pinuspollen season coincided with the coolestaverage monthly temperatures (< 22°C),lowest rainfall (< 7mm), and four weeks afterdaily minimum temperatures fell to 5–9°Cin late autumn. Correlations obtained betweendaily airborne Pinus pollen counts andtemperature/rainfall parameters show thatdensities of airborne Pinus pollen arenegatively correlated with maximum temperature(p < 0.0001), minimum temperature (p < 0.0001)and rainfall (p < 0.05) during the mainpollination period. The mean duration of eachpollen season was 52 days; longer seasons wereshown to be directly related to lower averageseasonal maximum temperatures (r² = 0.85,p = 0.025). These results signify that maximumand minimum temperatures are the majorparameters that influence the onset andduration of the Pinus pollen season inthe environs of Brisbane. Respiratory allergyis an important health issue in Brisbane,Australia, but it remains unknown whether ornot airborne Pinus pollen is acontributing factor.     

The EPSPS gene flow from glyphosate-resistant Brassica napus to untransgene B. napus and wild relative species Orychophragmus violaceus

Gene flow from transgenic plants to compatible wild relatives is one of the major impediments to the development of the culture of genetically engineered crop plants. In this work, the flow of EPSPS (conferring resistance to glyphosate) gene of transgene Brassica napus toward the untransgene B. napus and wild relative species Orychophragmus violaceus in an open field (1 ha) was studied. The data related to only the 2004 and 2005 autumn season on one location of southwest of China. Pollen dispersal and fertilization of the target plants were favored and a detailed analysis of the hybrid offspring was performed. In field, the data studied show that the gene flow frequency was 0.16% between GM and non-GM B. napus at a distance of 1 m from the transgenic donor area. The crosspollination frequency was 0.05% between GM and non-GM B. napus at a distance of 5 m from the transgenic donor area. At a distance of 10 m, no crosspollination was observed. According to the results of this study, B. napus transgene flow was low. However, the wild relative species O. violaceus could not be fertilized by the transgenic pollen of B. napus, no matter what the distance was.     

Comparison of some pollen concentrations in Finland and the Estonian SSR

Summary Pollen and spore concentrations were compared in Tartu, Estonia and in three sites in Finland (Turku, Kuopio and Oulu) during May–September 1989. The onset ofQuercus, Pinus, Poaceae,Urtica andRumex flowering started earlier in Tartu than at any site in Finland. The flowering ofJuniperus andArtemisia, on the other hand, began earlier in Turku and Kuopio than in Tartu.Pinus andJuniperus showed a significant correlation (number of pollen grains at the same date) between Tartu and Turku and between Turku and Kuopio. Poaceae andUrtica were correlated between all the sites, as wasRumex except between Tartu and Turku.Artemisia was correlated between Tartu and Turku, Turku and Oulu, and Kuopio and Oulu.Cladosporium correlated between Tartu and Turku. The pollen seasons of Poaceae,Urtica andRumex are prolonged towards the south.     

Transgene flow to hybrid rice and its male-sterile lines

Gene flow from genetically modified (GM) crops to the same species or wild relatives is a major concern in risk assessment. Transgenic rice with insect and/or disease resistance, herbicide, salt and/or drought tolerance and improved quality has been successfully developed. However, data on rice gene flow from environmental risk assessment studies are currently insufficient for the large-scale commercialization of GM rice. We have provided data on the gene flow frequency at 17 distances between a GM japonica line containing the bar gene as a pollen donor and two indica hybrid rice varieties and four male-sterile (ms) lines. The GM line was planted in a 640m² in an isolated experimental plot (2.4 ha), which simulates actual conditions of rice production with pollen competition. Results showed that: (1) under parallel plantation at the 0-m zone, the transgene flow frequency to the ms lines ranged from 3.145 to 36.116% and was significantly higher than that to hybrid rice cultivars (0.037–0.045%). (2) Gene flow frequency decreased as the distance increased, with a sharp cutoff point at about 1–2 m; (3) The maximum distance of transgene flow was 30–40 m to rice cultivars and 40–150 m to ms lines. We believe that these data will be useful for the risk assessment and management of transgenic rice lines, especially in Asia where 90% of world's rice is produced and hybrid rice varieties are extensively used. Shirong Jia, Feng Wang and Lei Shi contributed equally to this investigation.     

Analysis of airborne pollen fall in Sakarya, Turkey

In this study, pollen grains were identified using Durham sampler in the atmosphere of Sakarya in 2000 and 2001. During these two years, a total of 10 805 pollen grains were recorded. A total of 5 386 pollen grains per cm² were recorded in 2000 and a total of 5 419 pollen grains per cm² in 2001. Pollen fall in the years 2000–2001 comprised grains belonging to 40 taxa and some unidentified pollen grains. Of these taxa, 22 belonged to arboreal and 18 taxa to non arboreal plants. Total pollen grains consisted of 69.45% grains from arboreal plants, 28.11% grains from non-arboreal plants and 2.44% unidentified pollen grains. In the region investigated, Gramineae, Pinus sp., Quercus sp., Cupressaceae/Taxaceae, Salix sp., Platanus sp., Populus sp., Carpinus sp., Fagus sp., Chenopodiaceae/Amaranthaceae, Xanthium sp., Moraceae, Corylus sp., Fraxinus sp., and Urticaceae released the greatest amount of pollen. The season of maximum pollen fall was from March to May, with a prevalence of arboreal pollen in the first months, and of pollen from non-arboreal plants in the last months of the year.     

Pollen survival and spore dispersal on altitudinal clines: the case of the Popocatepetl volcano,Mexico

Pollen grains are abundant micro-organisms in the summer or dry season outdoor atmosphere. Moreover, they are fragile organisms, living for only a few days and their constant chemical interactions with their surrounding environment makes them prime candidates as biological indicators of ambient conditions. Volumetric samples were taken at different altitudes (3900–4400 m) on the Popocatepetl volcano (5452 m), 70 km south-east of Mexico City (2240 m). Results from September–October 1992 show that: (1) Local airborne pollen emission is negligible, most plant species found along the altitudinal transect being entomophilous. Indeed, maximum in vivo pollen viability was found at 4100 m a.s.l., where very few total airborne pollens were collected. (2) Regional pollen transport is found under 4000 m a.s.l., where pollen viability is discontinuous, being either below 50% or over 75%. (3) There is a long distance transport layer of pollen and spore dispersal below Mean Cloud Base, but over maximum wind speed, at a height of 4200–4300 m a.s.l., where pollen were found highly viable (>80%). (4) Pollen concentration or viability were not correlated with wind speed, relative humidity or temperature. However, the same factors were all significant regarding spore concentrations.     

Pollen movement within a continuous forest of wind-pollinated Araucaria angustifolia, inferred from paternity and TwoGener analysis

As a matter of fact, Araucaria angustifolia populations occur predominately in small and isolated stands; only a minor number of continuous natural forests of this dioecious wind-pollinated coniferous tree species remain. To implement reasonable conservation, breeding and restorations program it is necessary to have the knowledge of pollen dispersal distance and fine-scale genetic structure. In this paper, levels and dispersion distance of pollen and spatial genetic structure of A. angustifolia were investigated in a 14 ha transect in a continuous forest in Paraná State, Brazil. Analyses have been performed by the use of eight microsatellite loci, paternity and TwoGener approaches, and spatial autocorrelation analysis. In transect, 52 male and 56 female adult trees were mapped and genotyped, together with 190 seeds. In the present transect, A. angustifolia show spatial genetic structure at distances up to 75 m. Paternity analysis indicated that 54% of seeds were fertilized by pollen from trees outside the transect. The calculated average pollination distance within transect was 102 and 98 m based on the paternity analysis and TwoGener analysis, respectively. We found a significant pollen gene pool structure across seed-trees ( , P < 0.01) that corresponds to an effective number of pollen donors of 6.4 male trees or an effective pollination neighbourhood area (A _ep ) of 2.1 ha. The findings suggest long-distance pollen dispersion (>100 m) inside the continuous forest. However, the high proportion occurs in short-distance producing biparental and correlated mating as well as reducing the variance effective size.     

Microsatellite analysis of the maternal origin of <Emphasis Type="Italic">Myrica rubra</Emphasis> seeds in the feces of Japanese macaques

Identifying the maternal origin of dispersed seeds is a challenging task because it is impossible to directly track seed movement once an animal has ingested them. However, recent development of molecular techniques allows us to identify the maternal origin of dispersed seeds in natural plant populations. Here we analyzed the maternal origin of Myrica rubra seeds found in the feces of Yakushima macaques. We detected a high level of diversity among the dispersed seeds, with an average of 3.0 maternal origins (range 1–9) in each feces. The average dispersal distance was 270.0 m (range 20.4–634.0 m) and the average of distance between maternal trees of dispersed seeds in the same feces was 161.5 m (range 0–573.9 m). These results suggest that seed dispersal by macaques plays an important role in maintaining the genetic diversity of Myrica rubra populations.     

Performance of the Coriolis air sampler,a high-volume aerosol-collection system for quantification of airborne spores and pollen grains

The Coriolis δ air sampler manufactured by Bertin Technologies (France) is a continuous air sampler, dedicated to outdoor monitoring of airborne spores and pollen grains. This high-volume sampler is based on patented Coriolis technology delivering a liquid sample. The air is drawn into a conical vial in a whirling type motion using suction; particles are pulled against the wall by centrifugal force. Airborne particles are separated from the air and collected in a liquid medium. This innovative solution allows rapid analysis by several techniques including PCR assay and serological assay in order to measure the antigenicity/allergenicity of pollen grains and fungal spores. Also, traditional counting of pollen grains or taxa identification by optical microscopy can be done. A study has been carried out by the Health Protection Agency (HPA), Porton Down, UK, to measure the physical efficiency of the Coriolis air sampler. The physical efficiency of the sampler for collection of micro-organism-laden particles of various sizes has been compared with that of membrane filter samplers using the techniques described by ISO 14698-1. The Coriolis was operated simultaneously with membrane filter samplers in a controlled room where they were challenged with uniform-sized particles of different diameters containing bacterial spores. For the larger particle sizes, it was found that the physical efficiency of the Coriolis was 92% for 10-μm particles. The biological performance of the Coriolis in the collection of airborne fungal spores and pollen grains was evaluated in comparison with a Hirst spore trap (one-week tape-on-drum type sampler) which is one of the most frequently used traps in the measurement of outdoor pollen grain concentrations. The advantages and limitations of both technologies are discussed. The Coriolis was operated simultaneously with a Hirst spore trap in the sampling station of Réseau National de Surveillance Aérobiologique, France (RNSA); the pollen grain and fungal spore counts were analysed by optical microscopy. The pollen grain count m⁻³ collected was compared for both devices. The dispersion values were obtained and statistical analysis was carried out. This study shows that the Coriolis air sampler provided equivalent recovery of pollen grain and fungal spores compared with the volumetric trap standard method (not significantly different, W test, α = 0.05). Nowadays, the French-led project, acronym MONALISA, with financial support from the European Commission––Life-Environment (LIFE05 ENV/F/000068), is testing this innovative air sampler in order to measure the antigenicity/allergenicity of the main aeroallergen particles, i.e. Betula (birch), Poaceae (grasses), Parietaria (pellitory), Olea spp (olive tree), and Artemisia (mugwort) pollen grains, and Alternaria (fungal spores) to validate a new approach of monitoring instead of quantifying pollen grains by their morphology. The robustness and efficiency of the MONALISA system is being demonstrated at a national level throughout Europe in eight different countries with different bio-climatic and topography characteristics: France, UK, Finland, Poland, Spain, Portugal, Switzerland, and Italy.     

A satellite-based map of onset of birch (<Emphasis Type="Italic">Betula</Emphasis>) flowering in Norway

Birch (Betula pubescens L.) is by far the most common deciduous tree in Norway and birch forests define the forest line both northwards and upwards. Because of its mountainous topography, long fjords, and long length from north to south, Norway is climatically and ecologically very diverse. Therefore, developing pollen forecasts in Norway is a challenging task. In this study we use MODIS-NDVI (normalized difference vegetation index) satellite data with 250 m spatial resolution and 16-days time resolution for the period 2000–2007, and birch pollen counts from ten Burkad traps distributed throughout Norway, to characterize the onset of birch flowering in Norway. Four of the seven trap stations with long-term series show significant values at the 5% level or better between the MODIS-NDVI defined onset and the date when the annual accumulated birch pollen sum reaches 2.5% of the annual total. A map of Norway that shows the eight-year mean (2000–2007) onset of birch flowering was produced. It reveals large differences in the timing of the onset of birch flowering along the north–south and altitude gradients. The map provides useful general information that can be utilized by the Norwegian pollen forecast service. This study shows that remote sensing is a useful tool for not only characterizing the onset of the birch pollen season but also revealing regional differences not easily detected by pollen stations alone.     

<Emphasis Type="Italic">Hieracium</Emphasis> subgen. <Emphasis Type="Italic">Pilosella</Emphasis>: pollen stainability in sexual,apomictic and sterile plants

Two pollen stainability tests (Alexander’s stain and acetocarmine) were used to detect differences in pollen viability of the sexual, apomictic and sterile plants of Hieracium subgen. Pilosella. In sexual taxa (Hieracium bauhini and H. densiflorum), the average stainability was 93.7–98.4%. Similarly high stainability (92.2–97.2%) was found in the apomictic Hieracium pilosellinum and in the majority of the apomictic populations (or plants) of the pentaploid and hexaploid H. bauhini. In some apomictic plants of Hieracium bauhini the average pollen stainability was 49.0–75.4%. The lowest pollen stainability was found in the sterile plants, i.e. the triploid H. pistoriense (33.6%) and the pentaploid H. brachiatum (29.6%).     

Five decades of rapid forest spread in the Pieria Mountains (N. Greece) reconstructed by means of high-resolution pollen analysis and aerial photographs

The rapid spread of pine forests on the western side of the Pieria Mountains (N. Greece) began in 1945 and was facilitated by the recession of human activity and the siliceous geological substrate of the area. The pine expansion is visible not only in the vegetation maps for the years 1945, 1960 and 1992 but is also reflected in pollen data acquired from the high resolution analysis of a peat sequence covering the period 1945–1997, according to ²¹⁰Pb and ¹³⁷Cs dating. A possible inflation of the calculated PAR values due to discrepancies in the dating model is suspected and indirectly traced from the comparison of the Pinus mean PAR values for the most recent eight years of the monolith with those of a Tauber trap placed in the same mire. The presence of a high pollen producer such as pine enables the calculation of reliable PAR values, despite any discrepancies in the age-depth model. The Pinus PAR values “quantify” better the expansion of this timberline species, while pollen percentages reflect the same event in a “qualitative” way, i.e. as part of the general vegetation changes that take place in the area.     

Palm oil mill effluent (POME) cultured marine microalgae as supplementary diet for rotifer culture

Malaysia is the world’s leading producer of palm oil products that contribute US$ 7.5 billion in export revenues. Like any other agro-based industries, it generates waste that could be utilized as a source of organic nutrients for microalgae culture. Present investigation delves upon Isochrysis sp. culture in POME modified medium and its utilization as a supplement to Nanochloropsis sp. in rotifer cultures. The culture conditions were optimized using a 1 L photobioreactor (Temp: 23°C, illumination: 180 ∼ 200 μmol photons m⁻²s⁻¹, n = 6) and scaled up to 10 L outdoor system (Temp: 26–29°C, illumination: 50 ∼ 180 μmol photons m⁻²s⁻¹, n = 3). Algal growth rate in photobioreactor (μ = 0.0363 h⁻¹) was 55% higher compared to outdoor culture (μ = 0.0163 h⁻¹), but biomass production was 1.3 times higher in outdoor culture (Outdoor = 91.7 mg m⁻²d⁻¹; Photobioreactor = 69 mg m⁻²d⁻¹). Outdoor culture produced 18% higher lipid; while total fatty acids (FA) was not significantly affected by the change in culture systems as both cultures yield almost similar concentrations of fatty acids per gram of sample (photobioreactor = 119.17 mg g⁻¹; outdoor culture = 104.50 mg g⁻¹); however, outdoor cultured Isochrysis sp. had 26% more polyunsaturated fatty acids (PUFAs). Rotifers cultured in Isochrysis sp./ Nanochloropsis sp. (1:1, v/v) mixture gave similar growth rate as 100% Nanochoropsis sp. culture (μ = 0.40 d⁻¹), but had 45% higher counts of rotifers with eggs (t = 7, maximum). The Isochrysis sp. culture successfully lowered the nitrate (46%) and orthophosphate (83%) during outdoor culture.