Factors Influencing Allelopathy and Toxicity in Prymnesium parvum1

Granéli, Edna and Paulo S. Salomon, 2010. Factors Influencing Allelopathy and Toxicity in Prymnesium parvum. Journal of the American Water Resources Association (JAWRA) 46(1):108-120. Abstract: Some microalgae are able to kill or inhibit nutrient-competing microalgae, a process called allelopathy. Inhibiting or killing competitors enable these species to monopolize limiting resources, such as nitrogen and phosphorus. Prymnesium parvum is known to produce such allelopathic compounds, substances that seem identical to the ichthyotoxins identified from this species. Biotic and abiotic environmental factors influence not only growth rates but also toxin/allelopathic compounds production by P. parvum cells. Toxin production, as well as allelopathy, including grazer deterrence, increases dramatically in light, temperature, or nutrient stressed P. parvum cells. Correspondingly, toxicity and allelopathy may decrease, or cease completely, if cells are grown with high amounts of N and P in balanced proportions. However, even under nutrient (N and P) sufficient conditions, P. parvum is able to produce toxins/allelopathic compounds, with negative effects on other phytoplankton species or grazers, if cells densities of P. parvum are high relative to other species. This negative effect might shift the plankton community to more toxin resistant species. Filtrates from nutrient-deficient P. parvum cultures have almost the same strong negative effect on grazers and other phytoplankton species as when Prymnesium cells are grown together with the target organisms. Eutrophication, the increased input of N and P to aquatic ecosystems, besides increasing nutrient concentrations, is usually provoking unbalanced N:P condition for the optimal growth of phytoplankton, deviating from the Redfield ratio, i.e., the phytoplankton cellular nitrogen to phosphorus ratio, N:P = 16:1 (by atoms) or 7.2:1 (by weight). Eutrophication thus both enhances P. parvum growth and increases production of toxins and allelopathic compounds. Supplying N-deficient or P-deficient P. parvum cells with the deficient nutrient reduces toxicity to less than half within 24 h after additions. As P. parvum is mixotrophic, uptake of dissolved or particulate organic N (DON or PON) can also reduce toxicity and allelopathy in the same manner as addition of inorganic N to N-starved cells. In conclusion, P. parvum, by increasing its toxicity and allelopathic ability under poor environmental conditions, outcompetes the co-occurring phytoplankton species.     

Refining Nitrogen and Phosphorus Fertilization Strategies for Controlling the Toxigenic Alga Prymnesium parvum1

Kurten, Gerald L., Aaron Barkoh, Drew C. Begley, and Loraine T. Fries, 2010. Refining Nitrogen and Phosphorus Fertilization Strategies for Controlling the Toxigenic Alga Prymnesium parvum. Journal of the American Water Resources Association (JAWRA) 46(1):170-186. DOI: 10.1111/j.1752-1688.2009.00401.x Abstract: Previous studies have shown that three times weekly applications of phosphorus (30 μg P/l) and nitrogen (300 μg N/l) were effective at reducing the density and toxicity of the alga Prymnesium parvum in limnocorrals simulating a 40-day moronid (e.g., striped bass, Morone saxatilis, and palmetto bass, M. saxatilis ×Morone chrysops) fingerling culture period. However, this fertilization regime produced high pH and unionized ammonia-N concentrations that are detrimental to the survival of moronid fry and fingerlings. In two follow-up experiments we changed the source of N from ammonia to nitrate, reduced fertilization rates, and examined the effect of N-only or P-only fertilization. In the first experiment P fertilization rates were reduced by one-half to 15 μg P/l and NO₃-N was substituted for NH₃-N at the previously used rate of 300 μg N/l. In the second experiment, N fertilization rates were reduced to 150 μg N/l and the frequency of fertilization was determined by pH and P. parvum responses. Nitrate appeared to be as effective as ammonia as a source of N and when used in combination with P reduced P. parvum cell density and ichthyotoxicity. However, reduced N and P application rates and lower pond water temperatures during the study appeared to have decreased the speed at which fertilization produced these effects. While lower fertilization rates reduced algal productivity, high pH remained a concern for fish culture although pH was reduced to levels that might be acceptable with careful management of fish culture activities. Neither N-only nor P-only fertilization had a measurable effect on algal productivity or eliminated P. parvum and its toxicity. Furthermore, P-only fertilization may have increased P. parvum density and toxicity. For controlling P. parvum density and ichthyotoxicity we recommend a fertilization rate of 212 μg NO₃-N/l plus 30 μg PO₄-P/l applied three times weekly for aquaculture ponds where high pH is not a concern. Where high pH is a concern we recommend a fertilization rate of 117 μg NO₃-N/l plus 16 μg PO₄-P/l applied three times weekly with careful attention to afternoon pond pH.     

Current Status of Mathematical Models for Population Dynamics of Prymnesium parvum in a Texas Reservoir1

Grover, James P., Jason W. Baker, Daniel L. Roelke, and Bryan W. Brooks, 2010. Current Status of Mathematical Models for Population Dynamics of Prymnesium parvum in a Texas Reservoir. Journal of the American Water Resources Association (JAWRA) 46(1):92-107. DOI: 10.1111/j.1752-1688.2009.00393.x Abstract: Blooms of the harmful alga Prymnesium parvum have apparently increased in frequency in inland waters of the United States, especially in western Texas. A suite of mathematical models was developed based on a chemostat (or continuously stirred tank reactor) framework, and calibrated with data from Lake Granbury, Texas. Inputs included data on flows, salinity, irradiance, temperature, zooplankton grazing, and nutrients. Parameterization incorporated recent laboratory studies relating the specific growth rate of P. parvum to such factors. Models differed in the number of algal populations competing with P. parvum, and whether competition occurred only by consumption of shared nutrients, or additionally through production of an allelopathic chemical by one of the populations, parameterized as cyanobacteria. Uncalibrated models did not reproduce the observed seasonal dynamics of P. parvum in Lake Granbury, which displayed a maximum population in late February during a prolonged bloom in cooler weather, and reduced abundance in summer. Sensitivity analyses suggested two modifications leading to predictions that better resembled observations. The first modification greatly reduces the optimal temperature for growth of P. parvum, an approach that disagrees with laboratory experiments indicating a strong potential for growth at temperatures above 20°C. The second modification increases the growth rate of P. parvum at all temperatures, in models including cyanobacterial allelopathy. Despite these adjustments, calibrated models did not faithfully simulate all features of the seasonal dynamics of P. parvum.     

Hemolysis,Fish Mortality,and LC-ESI-MS of Cultured Crude and Fractionated Golden Alga (Prymnesium parvum)1

Schug, Kevin A., Theodore R. Skingel, Sandra E. Spencer, Carlos A. Serrano, Cuong Q. Le, Christopher A. Schug, Theodore W. Valenti, Jr., Bryan W. Brooks, Laura D. Mydlarz, and James P. Grover, 2010. Hemolysis, Fish Mortality, and LC-ESI-MS of Cultured Crude and Fractionated Golden Alga (Prymnesium parvum). Journal of the American Water Resources Association (JAWRA) 46(1):33-44. DOI: 10.1111/j.1752-1688.2009.00389.x Abstract: Erythrocyte lysis and fish mortality assays, in combination with high performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis, were investigated for bioassay-guided fractionation of cultured golden alga (Prymnesium parvum). Intracellular constituents from isolated cell pellets and extracellular supernatant growth medium were fractionated by a variety of common separation modes, including reversed phase and normal phase solid phase extraction step fractionation procedures. For reversed phase fractionation of extracellular growth medium, one fraction was obtained that displayed hemolytic activity and adversely affected fish survival. Effective dose concentrations for this sample were similar in both assays and the LC-ESI-MS analysis of the fraction showed a number of mass spectral signals which were distinct to this fraction. Fractions obtained from separation of an ethanol extract of the lyophilized cell pellet provided one sample that was highly hemolytic, but not toxic to fish. Discrepancies such as this, along with notable fish behavioral responses from other nonhemolytic cell pellet fractions, problems with the use of unbonded silica gel for fractionation, and misleading mass spectral signatures are interesting in the context of our current understanding of P. parvum toxicity and remain to be investigated further. This work provides an account of ongoing research aimed toward comprehensive elucidation of toxic constituents produced by golden alga for the purpose of providing a better understanding and means to potentially remediate the ecological impact of this harmful bloom organism.     

Prymnesium parvum: The Texas Experience1

Southard, Gregory M., Loraine T. Fries, and Aaron Barkoh, 2010. Prymnesium parvum: The Texas Experience. Journal of the American Water Resources Association (JAWRA) 46(1):14-23. DOI: 10.1111/j.1752-1688.2009.00387.x Abstract: Golden alga Prymnesium parvum was first identified in Texas during a fish kill investigation on the Pecos River in 1985. Since then golden alga kills occurred sporadically in a variety of waters in the western part of the state until 2001 when the alga became endemic in the Brazos, Canadian, Colorado, Red, and Rio Grande river systems, including the water supplies of two public fish hatcheries, the Possum Kingdom and Dundee state fish hatcheries. The increasing area adversely affected by the alga and frequent massive fish kills heightened public and political awareness and concerns regarding the ecological and economic impacts of P. parvum blooms. The Texas Parks and Wildlife Department (TPWD), the wildlife conservation agency of the state, responded to these concerns with a program to assess the ecological and economic impacts and to develop management options. To date 33 water bodies have been affected and losses are conservatively estimated at 34 million fish valued at US$13 million. Several sport fisheries, including smallmouth bass Micropterus dolomieu, striped bass Morone saxatilis, channel catfish Ictalurus punctatus, and blue catfish Ictalurus furcatus, have been severely affected. Additionally, 26 imperiled fish species occur in the affected water basins and some have been adversely affected. Economic losses associated with reduced fishing and other water-based recreational activities appear considerable. The combined economic losses to three counties (Palo Pinto, Stephens, and Young) surrounding Possum Kingdom reservoir for 2001 and 2003 were estimated at US$2.8 million and US$1.1 million, respectively. This paper describes how the TPWD responded to public and political concerns relative to the emergence of golden alga, its harmful effects to fisheries, and its historic and current statewide distribution.     

Prymnesium parvum Population Dynamics During Bloom Development: A Role Assessment of Grazers and Virus1

Schwierzke, Leslie, Daniel L. Roelke, Bryan W. Brooks, James P. Grover, Theodore W. Valenti, Jr., Mieke Lahousse, Carrie J. Miller, and James L. Pinckney, 2010. Prymnesium parvum Population Dynamics During Bloom Development: A Role Assessment of Grazers and Virus. Journal of the American Water Resources Association (JAWRA) 46(1):63-75. DOI: 10.1111/j.1752-1688.2009.00391.x Abstract: The toxic haptophyte Prymnesium parvum is a harmful alga known to cause fish-killing blooms that occur worldwide. In Texas (United States), P. parvum blooms occur in inland brackish water bodies and have increased in frequency and magnitude in recent years. In this study we conducted three consecutive field experiments (Lake Whitney) to investigate the influence of zooplankton and viruses on P. parvum bloom dynamics during the time of year when blooms are still typically active in Texas (early spring). A localized P. parvum bloom developed during our study that involved increasing levels of toxicity (based on Pimephales promelas and Daphnia magna bioassays). Only in our last experiment, during later stages of bloom development and under highly toxic conditions, did the presence of grazers show a statistically significant, negative effect on P. parvum population dynamics. During this experiment, a rotifer-dominated zooplankton community emerged, composed mostly of Notholca laurentiae, suggesting that this species was less sensitive than other grazers to toxins produced by P. parvum. Microzooplankton may have also been important at this time. Similarly, only our final experiment demonstrated a statistically significant, negative effect of viruses on P. parvum. This exploratory study, resulting in observed impacts on P. parvum populations by both grazers and virus, enhances our understanding of P. parvum ecology and highlights direction for future studies on resistance of zooplankton to prymnesin toxins and algal-virus interactions.     

Influence of Prey and Nutritional Status on the Rate of Nitrogen Uptake by Prymnesium parvum (haptophyte)1

Lindehoff, Elin, Edna Granéli, and Patricia M. Glibert, 2010. Influence of Prey and Nutritional Status on the Rate of Nitrogen Uptake by Prymnesium parvum (Haptophyte). Journal of the American Water Resources Association (JAWRA) 46(1):121-132. DOI: 10.1111/j.1752-1688.2009.00396.x Abstract: We studied how the specific nitrogen (N) uptake rates of nitrate (NO₃⁻), urea, and the amino acids, glutamic acid and glycine, by Prymnesium parvum were affected by (1) the change from N-deficient status to N-sufficient status of the P. parvum cells, (2) presence of prey from a natural Baltic Sea plankton community, and (3) the composition of prey as affected by additions of terrestrial originated dissolved organic matter (DOM) or inorganic nutrients. Nitrogen-deficient P. parvum (16 μM NO₃⁻ and 4 μM PO₄⁻, molar N:P ratio of 4:1) were mixed with a natural Baltic plankton community and given PO₄³⁻ and (1) NO₃⁻ (control) or (2) high molecular weight DOM, >1 kDa concentrated from sewage effluent (+DOM), in a molar N:P ratio of 9-10:1. With additions of ¹⁵N-enriched substrates, rates of N uptake from NO₃⁻, urea, and the amino acids glycine and glutamic acid were measured every 24 h for 72 h. Initial N-deficient P. parvum were highly toxic (3.7 ± 0.9 × 10⁻⁴ mg Sap equiv/cell) and toxic allelochemicals were released into the medium causing the natural plankton community to lyse. Rates of N uptake differed between the “control” and the “+DOM” treatments over time; total (sum of the N substrates measured) absolute uptake rates (ρ_cell, fmol N/cell/h) at ambient culture conditions were significantly higher (ANOVA, p < 0.05) in the more toxic “control” treatments compared with the “+DOM” treatments after 48 h. In the “control” treatment, the total ρ_cell increased significantly (ANOVA, p < 0.01) from time 0 to 48 h, while in the “+DOM” treatment there was no significant increase. Released organic nutrients from the lysed plankton cells may have increased uptake rates of amino acids and urea by P. parvum. All uptake rates declined in all treatments by 72 h. Total dissolved N uptake rates at ambient culture conditions were estimated to make up about 10% of the N P. parvum are potentially capable of ingesting from particulate prey.     

Prymnesium parvum: The Norwegian Experience1

Johnsen, Torbjørn M., Wenche Eikrem, Christine D. Olseng, Knut E. Tollefsen, and Vilhelm Bjerknes, 2010. Prymnesium parvum: The Norwegian Experience. Journal of the American Water Resources Association (JAWRA) 46(1):6-13. DOI: 10.1111/j.1752-1688.2009.00386.x Abstract: In Norwegian waters, Prymnesium parvum has been reported from Oslofjorden in the south to Spitzbergen in the north. However, blooms of P. parvum have only been reported from the Sandsfjorden system in Ryfylke, Western Norway where the salinity of the permanent brackish layer (2-5 m) typically is in the range of 4-7 psu during the summer months. The first bloom on record occurred in 1989, and it killed 750 metric tons of caged salmon and trout which was a significant economic loss to the fish farming industry. Toxic blooms occurred as well in subsequent years and the number of fish farms in the area decreased considerably as did the occurrence of P. parvum. In 2005, fish farming was reintroduced to the area and again, in 2007 a toxic bloom of P. parvum killed 135 metric tons of caged fish. The Norwegian Institute for Water Research has, in collaboration with “Erfjord Stamfisk” fish farm, set up a monitoring program that includes light microscopy cell counts of Prymnesium, water quality measurements, and observation of the caged fish. A submergible fish net was mounted over the fish pens and during the toxic outbreak of P. parvum in July-August 2007, which was as previous years confined to the upper brackish water layer, the fish nets were lowered to 10 m depths below the surface and fish feeding was temporarily stopped. Despite substantial weight loss, the fish survived the toxic bloom and the economic loss was minimal. Monitoring of P. parvum bloom dynamics in 2007 revealed that populations were initially dominated by the nonmotile forms which were gradually replaced by the flagellated forms. Toxicity was observed when the flagellated cells dominated populations in the summer. Chrysochromulina, solitary small Chaetoceros species, and small centric diatoms co-existed with P. parvum during the monitoring period (June-October).     

Prymnesium parvum Control Treatments For Fish Hatcheries1

Barkoh, Aaron, Dennis G. Smith, and Gregory M. Southard, 2010. Prymnesium parvum Control Treatments for Fish Hatcheries. Journal of the American Water Resources Association (JAWRA) 46(1):161-169. DOI: 10.1111/j.1752-1688.2009.00400.x Abstract: In 2001, the ichthyotoxic microalga Prymnesium parvum caused massive fish kills and adversely affected fish production at the Texas Parks and Wildlife Department (TPWD) Dundee State Fish Hatchery. Since then, we have investigated several P. parvum bloom and ichthyotoxicity control treatments to develop management strategies that allow fish production and prevent the spread of the alga into unaffected hatcheries and impoundments. Current control successes include treatments for ponds, water supply, and a hazard analysis and critical control point program. For pond treatment, ammonium sulfate (as 0.14-0.25 mg/l un-ionized ammonia nitrogen for temperatures above 15°C), copper sulfate (2 mg/l), Cutrine^®-Plus (0.2-0.4 mg/l as copper), or potassium permanganate (3 mg/l above the potassium permanganate demand) controls P. parvum blooms. Copper sulfate at 1 mg/l controls P. parvum but is unable to eliminate ichthyotoxicity whereas potassium permanganate at 2 mg/l above the potassium permanganate demand controls ichthyotoxicity. For water treatment, ultraviolet (UV) light at 193-220 mJ/cm² doses or ozone at 0.4-1.2 mg/l for 6 min destroy P. parvum cells and reduce or eliminate ichthyotoxicity. A combination UV and ozone treatment appears to provide the best results; however, successful treatments depend on dosage relative to cell density and toxin concentration. To prevent the spread of the alga, hatchery fish delivery units and equipment are cleaned with household bleach (10% solution for 15 minutes) or hydrogen peroxide (62.5-12,500 mg/l for 0.25-24 hours). These treatments are tailored to water quality conditions and the fish species cultured at affected TPWD hatcheries. We recommend that other users test these treatments before applying them to ponds or other impoundments containing fish or other aquatic life.     

Factors Influencing Prymnesium parvum Population Dynamics During Bloom Initiation: Results from In-lake Mesocosm Experiments1

Roelke, Daniel L., Leslie Schwierzke, Bryan W. Brooks, James P. Grover, Reagan M. Errera, Theodore W. Valenti, Jr., and James L. Pinckney, 2010. Factors Influencing Prymnesium parvum Population Dynamics During Bloom Initiation: Results from In-Lake Mesocosm Experiments. Journal of the American Water Resources Association (JAWRA) 46(1):76-91. DOI: 10.1111/j.1752-1688.2009.00392.x Abstract: The alga Prymnesium parvum forms large fish-killing blooms in many Texas lakes. In some of these lakes, however, P. parvum occurs but does not develop blooms. In this study, we investigated factors that may influence bloom initiation by conducting a series of in-lake experiments involving mixing of waters from Lake Whitney, which has a history of P. parvum blooms, with waters from Lake Waco where no blooms have occurred. In all experiments, the addition of Lake Waco waters resulted in a poorer performance of P. parvum. Various experimental treatments and field data show that differences in grazing, pathogens, nutrients, and salts between the two lakes were not likely factors that contributed to this observation. Industrial and agricultural contaminants, allelochemicals and algicidal chemicals were not measured as a part of this research. However, anthropogenic contaminants other than nutrients were not observed at levels exceeding water quality standards in Lake Waco in recent years. On the other hand, nuisance cyanobacteria are common in Lake Waco, where Microcystis sp. and Anabaena sp. were abundant during the initiation of our experiments, both taxa are known to produce chemicals with allelopathic properties. In addition, the emergent field of algal-heterotrophic bacteria interactions suggests that chemicals produced by heterotrophic bacteria should not be overlooked. Further research focusing on the chemical interactions between cyanobacteria and P. parvum, as well as the potential role of algicidal bacteria, in the initiation of P. parvum blooms is necessary, as it may be important to the management of these blooms.     

Comparison of Three Algaecides for Controlling the Density of Prymnesium parvum1

Rodgers, John H., Jr., Brenda M. Johnson, and West M. Bishop, 2010. Comparison of Three Algaecides for Controlling the Density of Prymnesium parvum. Journal of the American Water Resources Association (JAWRA) 46(1):153-160. DOI: 10.1111/j.1752-1688.2009.00399.x Abstract: Prymnesium parvum has become more prevalent in water resources of the southern United States. As the potential impacts of P. parvum are relatively well known, especially its capability to severely affect fish, managers have sought efficacious, environmentally sound, and socially acceptable strategies for mitigating this noxious species. Laboratory testing was used to identify an effective algaecide for control of P. parvum from Texas, Arizona, Florida, North Carolina, and South Carolina. Cutrine^®-Plus at 0.2 mg Cu/l significantly decreased the density of P. parvum in samples from all of these locations. Both copper sulfate and Phycomycin^® were less effective for controlling the population growth of P. parvum. The predicted response from the laboratory study was confirmed in the field at the Arizona site. Strategic use of Cutrine^®-Plus in larger water resources could provide toxin-free refugia to allow some fish to survive and repopulate the water resource when the golden alga infestation abates.     

Potential Methods for Managing Prymnesium parvum Blooms and Toxicity,With Emphasis on Clay and Barley Straw: A Review1

Hagström, Johannes A., Mario R. Sengco, and Tracy A. Villareal, 2010. Potential Methods for Managing Prymnesium parvum Blooms and Toxicity, With Emphasis on Clay and Barley Straw: A Review. Journal of the American Water Resources Association (JAWRA) 46(1):187-198. DOI: 10.1111/j.1752-1688.2009.00402.x Abstract: Harmful algal bloom (HAB) control and mitigation is a complex problem in ecosystem management. Phytoplankton play an important role in aquatic ecosystems as primary producers and food sources for many commercially important shellfish and there are limited options for targeting just a single species within the community. Chemical treatments (e.g., algaecides), rotting barley straw, nitrogen and phosphorus manipulation, and clay and/or flocculants are but a few techniques tested or used to reduce fish kills or shellfish contamination during a HAB event. Prymnesium parvum control has focused on the use of chemicals, nutrient manipulation, and clay flocculation. However, many HAB control methods have been rejected due to their effects on ecosystems, high costs, or limited effects on target organisms. For example, rotting barley straw (Hordeum vulgare) is considered to be an environmentally friendly alternative, but has been found to have very different results on the phytoplankton community depending on the dominating taxa and is ineffective against P. parvum and dinoflagellate blooms. Clay flocculation is a useful control/mitigation technique during fish kills in marine aquaculture sites in South Korea and can be effective in freshwater if the correct combination of clay and flocculent is used. Toxins produced by P. parvum and Karenia brevis also bind to phosphatic clay, thereby removing and/or neutralizing the toxins, but there is concern that the clay will have a negative effect on sessile organisms. Some shellfish suffer high mortalities and significant impacts on somatic and reproductive tissue growth at high clay loads; however, benthic communities appear to be unchanged after five years of clay treatment in South Korea. There are likely site-specific and ecosystem-specific characteristics that make generalizations about control options difficult and require careful assessment of options at each location.     

Combining Flow Cytometry and Real-Time PCR Methodology to Demonstrate Consumption by Prymnesium parvum1

Bowers, Holly A., Andreas Brutemark, Wanderson F. Carvalho, and Edna Granéli, 2010. Combining Flow Cytometry and Real-Time PCR Methodology to Demonstrate Consumption by Prymnesium parvum. Journal of the American Water Resources Association (JAWRA) 46(1):133-143. DOI: 10.1111/j.1752-1688.2009.00397.x Abstract: Harmful algal bloom species can persist in the environment, impacting aquatic life and human health. One of the mechanisms by which some harmful algal bloom species are able to persist is by consumption of organic particles. Methods to demonstrate and measure consumption can yield insight into how populations thrive. Here, we combine flow cytometry and real-time PCR to demonstrate consumption of a cryptophyte species (Rhodomonas sp.) by a toxic mixotrophic haptophyte (Prymnesium parvum). Using flow cytometry, the feeding frequency of a population of P. parvum cells was calculated using the phycoerythrin (PE) fluorescence signal from Rhodomonas sp. and the fluorescence of an acidotropic probe labeling the food vacuoles. Feeding frequency increased in the beginning of the experiment and then began to decline, reaching a maximum of 47.5% of the whole P. parvum population after 212 min. The maximum number of consumed Rhodomonas sp. cells was 0.8 per P. parvum cell, and occurred after 114 min corresponding to an ingestion rate of 0.4 Rhodomonas sp. cells/P. parvum/h. Cells from the feeding P. parvum population were sorted, washed, and subjected to a real-time PCR assay targeting the cryptophyte 18S locus. There was a correlation between cycle threshold (Ct) values and number of consumed prey cells calculated by fluorescence. Overall, this study shows that flow cytometric analysis, of the acidotropic probe and prey pigments, is an efficient and rapid tool in enumerating food vacuoles and the number of prey cells consumed. Furthermore, we suggest that real-time PCR can be applied to cells sorted by flow cytometry, thus allowing for the detection and potential quantification of the targeted prey cells.     

Mini Review: Probes for Detecting Prymnesium parvum and Preliminary Results From Gene Expression Studies1

Medlin, Linda K., Sonja Diercks, and Sara Beszteri, 2010. Mini Review: Probes for Detecting Prymnesium parvum and Preliminary Results From Gene Expression Studies. Journal of the American Water Resources Association (JAWRA) 46(1):144-152. DOI: 10.1111/j.1752-1688.2009.00398.x Abstract: Prymnesium parvum is common in brackish and marine coastal waters within temperate zones, world-wide. P. parvum forms recurrent blooms causing fish kills in many parts of the world. Harmful blooms are formed in nutrient rich, low salinity lakes, ponds, river systems, or estuaries. Probes made to this species and to the genus Prymnesium have been tested in dot blot and fluorescent in situ hybridization (FISH) and used with a solid-phase cytometer using a tyramide signal amplification (TSA) enhanced FISH hybridization to provide a means to identify the cells before blooms develop with automated counting. Field counts from light microscopy have been compared with solid-phase cytometer counts. Additional detection systems, such as biosensors and microarrays, have also been developed to identify rapidly this species without resorting to electron microscopy. Advantages and disadvantages of each detection system are discussed. A study of the genes expressed by P. parvum under environmental conditions that can induce blooms or stress was undertaken to try to understand the ecology behind toxic blooms.     

Global Genetic Relationships Among Isolates of Golden Alga (Prymnesium parvum)1

Lutz-Carrillo, Dijar J., Gregory M. Southard, and Loraine T. Fries, 2010. Global Genetic Relationships Among Isolates of Golden Alga (Prymnesium parvum). Journal of the American Water Resources Association (JAWRA) 46(1):24-32. DOI: 10.1111/j.1752-1688.2009.00388.x Abstract: Prymnesium parvum is considered among the most harmful algal species in the world for finfish and other gill breathing organisms. Although it is globally distributed, with the exception of Antarctica, P. parvum is usually associated with coastal and brackish waters. Historically, P. parvum incidents were recorded in the eastern hemisphere; however, in 1985 it was detected in inland Texas waters. We used DNA sequence variation of the first internal transcribed spacer in the nuclear ribosomal operon (ITS1) among multiple samples of P. parvum from Texas and other locales to address the possible origins of P. parvum in Texas and the United States (U.S.). With the exception of a sample from Diversion Lake, other samples from Texas, South Carolina, and Wyoming exhibited limited genetic variation and were similar in sequence to a sample from Scotland. The Diversion Lake sample was similar in sequence to samples from Denmark and Norway, and the Maine sample was highly similar to samples from England. These results suggest multiple independent introductions of P. parvum to the U.S.     

STORM AND SEASONAL DISTRIBUTIONS OF FECAL COLIFORMS AND CRYPTOSPORIDIUM IN A SPRING1

ABSTRACT: The transmission of disease in ground water is a topic of great concern to government agencies, ground water specialists, and the general public. The purpose of this study was to compare the temporal variability in storm flow of fecal coliform bacteria densities and Cryptosporidium parvum oocyst densities in agriculturally impacted karst ground water. Cryptosporidium parvum oocyst densities ranged from 0 to 1,050 oocysts/1, and mean storm densities ranged from 3.5 to 156.8 oocysts/1. Fecal coliform densities ranged from less than 1 CFU/100ml to more than 40,000 CFU/100ml, and geometric mean storm densities ranged from 1.7 CFU/100ml to more than 7,000 CFU/100ml. Fecal coliform densities correlated well with flow during storms, but Cryptosporidium oocyst densities exhibited a great deal of sample to sample variability and were not correlated with flow. Fecal coliform densities did not correlate positively with Cryptosporidium oocyst densities. Fecal coliform densities were greatest at storm peaks, when sediment loads were also greatest. Multiple transport mechanisms for fecal coliform bacteria and C. parvum oocysts may necessitate various agricultural land management and livestock health maintenance practices to control movement of pathogens to karst ground water.     

Biohythane production from pineapple peel using Methanosarcina mazei enhanced with palm oil mill effluent (POME) sludge in single-stage anaerobic digestion

Biohythane production via single-stage anaerobic digestion (AD) is an effective way for sustainable energy recovery from lignocellulosic biomass. In this paper, biohythane was produced through the AD process from pineapple peel waste substrate using purely cultured Methanosarcina mazei with the enhancement of palm oil mill effluent (POME) sludge as the inoculum. This study focuses on the effects of the lignocellulosic pre-treatment method, the addition of POME sludge into M. mazei culture medium as inoculum, and various operational conditions (food to microorganisms (F/M) ratios, temperature, pH) on gas production performances. The experimental results indicate that these parameters influenced the efficiency of biohythane production by producing the peak maximum biohythane production rate values (HPR_max) and (MPR_max), H₂:CH₄ = 1.93:0.67 L/L-d, and biohythane yield (HY) and (MY), H₂:CH₄ = 1.18:0.55 mL/L-substrate. This study demonstrates that biohythane gas (H₂ + CH₄ + CO₂) production from pineapple waste can be accelerated by M. mazei only with the enhancement of POME sludge through single-stage AD system under mesophilic batch process conditions.     

Changing effluent chemistry affect survival,growth and physiological function of <Emphasis Type="Italic">Acacia nilotica</Emphasis> seedlings in northwestern region of India

Recycling and conservation efforts for water are the need of the day because of the lack of new water sources and the ever-increasing demand for drinking water. Seedlings of Acacia nilotica L. were irrigated with: canal water (T₁, control); municipal effluent (T₂); textile effluent (T₃); steel effluent (T₄); textile + municipal effluent in 1:1 ratio (T₅); steel + municipal effluent in 1:2 ratio (T₆); steel + textile in 1:2 ratio (T₈) and steel + municipal + textile in 1:2:2 ratio (T₇) with views to observe effluents effect on the seedlings and its adaptability and to recommend safe disposal of these effluents. Seedlings in T₆, T₇ and T₈ showed 50% lesser height and collar diameter than those in control. Seedlings in T₂ attained greatest height, collar diameter, numbers of branches and produced 140 g dry biomass seedling⁻¹. Highest concentration of manganese (Mn), iron (Fe), copper (Cu) and zinc (Zn) and lowest concentration of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) in the seedlings of T₄, T₆, T₇ and T₈ resulted in nutritional imbalance, mineral toxicity and reduction in photosynthetic (Pn) and transpiration (E) rates and caused seedling mortality. Seedlings of T₃ had highest sodium concentration and low concentration of Ca, Mg and micronutrients resulting in nutritional imbalance, augmented chlorosis and reduced gas exchange and biomass by half as compared to control. Increased growth, Pn and E and biomass in seedlings of T₅ over T₃ and survival period in T₆, T₇ and T₈ seedlings suggested a beneficial effect of effluents mixing. Unscientific disposal should be avoided and toxic concentration of metal ions␣may be reduced for long-term application and harmless disposal of effluents in afforestation and urban development.     

Benthic Algal (Periphyton) Growth Rates in Response to Nitrogen and Phosphorus: Parameter Estimation for Water Quality Models

Nitrogen (N) and phosphorus (P) are significant pollutants that can stimulate nuisance blooms of algae. Water quality models (e.g., Water Quality Simulation Program, CE‐QUAL‐R1, CE‐QUAL‐ICM, QUAL2k) are valuable and widely used management tools for algal accrual due to excess nutrients in the presence of other limiting factors. These models utilize the Monod and Droop equations to associate algal growth rate with dissolved nutrient concentration and intracellular nutrient content. Having accurate parameter values is essential to model performance; however, published values for model parameterization are limited, particularly for benthic (periphyton) algae. We conducted a 10‐day mesocosm experiment and measured diatom‐dominated periphyton biomass accrual through time as chlorophyll a (chl a) and ash‐free dry mass (AFDM) in response to additions of N (range 5–11,995 µg nitrate as nitrogen [NO₃‐N]/L) and P (range 0.89–59.51 µg soluble reactive phosphorus/L). Resulting half‐saturation coefficients and growth rates are similar to other published values, but minimum nutrient quotas are higher than those previously reported. Saturation concentration for N ranged from 150 to 2,450 µg NO₃‐N/L based on chl a and from 8.5 to 60 µg NO₃‐N/L when based on AFDM. Similarly, the saturation concentration for P ranged from 12 to 29 µg‐P/L based on chl a, and from 2.5 to 6.1 µg‐P/L based on AFDM. These saturation concentrations provide an upper limit for streams where diatom growth can be expected to respond to nutrient levels and a benchmark for reducing nutrient concentrations to a point where benthic algal growth will be limited.     

Polyurethane rotating disc system for post-treatment of anaerobically pre-treated sewage

The performance of polyurethane rotating discs (RBC-1) versus polystyrene rotating discs (RBC-2) for the treatment of an up-flow anaerobic sludge blanket (UASB) reactor effluent fed with domestic wastewater was investigated. Both RBC units were operated at the same organic loading rate (OLR) of 10.5 gCOD/m² d. and a hydraulic retention time (HRT) of 2.5 h. The residual values of COD fractions (COD_suspended, COD_colloidal and COD_soluble) in the treated effluent of RBC-1 and RBC-2 were similar. However, the removal efficiency of ammonia in the RBC-1 (87 ± 4%) was significantly higher than that found for RBC-2 i.e. 24 ± 6%. Moreover, RBC-1 achieved a substantial removal efficiency of 99.0 ± 1% for Escherichia coli (E. coli), while RBC-2 removed 91.2 ± 0.3%. Based on these results, optimization of RBC-1 treating UASB reactor effluent was extensively performed. The RBC-1 was operated at an OLR's of 4.0, 11 and 23 gCOD/m² d. The results obtained showed that increasing the OLR from 11.0 to 23.0 gCOD/m² d and decreasing the HRT from 2.5 to 1.25 h significantly declined the effluent quality of COD_total and ammonia. However, the residual values of COD_total and ammonia remained unaffected when increasing the OLR from 4.0 to 11.0 gCOD/m² d and by decreasing the HRT from 5 to 2.5 h. Bacteriological examination showed that the mean residual count of E. coli remained at a level of 10⁴/100 ml, in the effluent of RBC-1 independent on the imposed HRT. Accordingly, it is recommended to operate RBC-1 for treatment of anaerobically pre-treated sewage at an OLR of 11 gCOD/m² d and an HRT of 2.5 h.A feed-less (ammonia limitation) period of 9.0 days followed by 9.0 days feeding with high OLR of 26 gCOD/m² d. (raw sewage) was investigated to elaborate, if the nitrifiers of the RBC-1 are capable to convert ammonia to nitrate after totally 18 days when retuning back to the normal operating conditions. The results of the experiment clearly show a strong and immediate detrimental effect of imposing high OLR of 26 gCOD/m² d on the nitrification process in the nitrifying RBC unit. However, after returning back to the original OLR of 10.6 gCOD/m² d, the nitrification efficiency in the RBC unit was recovered within 2–3 days.