Free Ammonia Inhibition of Algal Photosynthesis in Intensive Cultures

The effect of free NH₃ inhibition on short-term photosynthesis was investigated in three microalgal species: the freshwater chlorophyte Scenedesmus obliquus, the marine diatom Phaeodactylum tricornutum and the marine chlorophyte Dunaliella tertiolecta. By performing a series of assays at various concentrations of added NH₄Cl and culture pH, we demonstrated that the inhibitory compound was free NH₃ and that pH played no role in determining the magnitude of inhibition, other than in establishing the degree of dissociation of nontoxic NH₄⁺ to toxic NH₃. When corrections were made for pH, all three species displayed the same sigmoidal response curve to free NH₃ concentration; 1.2 mM NH₃ led to 50% reduction in photoassimilation of ¹⁴C. Based on literature values, some marine phytoplankton appear to be significantly more sensitive to free NH₃ than were the test species, which are noted for their excellent growth characteristics. However, the combination of low algal biomass and strong pH buffering commonly found in most marine and many freshwater environments probably limits the possibilities for NH₃ toxicity to low alkalinity freshwaters and intensive algal cultures in which NH₄⁺ is the main source of N. Such conditions occur commonly in algal wastewater treatment systems.     

Carbon limitation of biomass production in high-rate oxidation ponds

Theoretical considerations confirmed by outdoor experiments indicated carbon limitation of biomass production in high-rate oxidation ponds at certain seasonal and operational conditions. Apparently, free carbon dioxide concentration in the pond is the major determinant of carbonlimiting algal photosynthesis. High concentrations of free CO(2) are provided through bacterial respiration which is the main contributor to algal photosynthesis. At high photosynthetic activities and low organic loadings, free CO(2) concentrations are low; its flux into algal cells determines photosynthesis and biomass production rate in the pond.     

Toxicity of ammonia to algae in sewage oxidation ponds.

Ammonia, at concentrations over 2.0 mM and at pH values over 8.0, inhibits photosynthesis and growth of Scenedesmus obliquus, a dominant species in high-rate sewage oxidation ponds. Photosynthesis of Chlorella pyrenoidosa, Anacystis nidulans, and Plectonema boryanum is also susceptible to ammonia inhibition. Dark respiration and cell morphology were unaffected by any combination of pH and ammonia concentrations tested, thus limiting the apparent effect to inhibition of the normal function of the chloroplasts. Methylamine had the same effect as ammonia, and its penetration into the cells was found to be pH dependent. Therefore, the dependence of toxicity of amines to algae on pH apparently results from the inability to penetrate the cell membrane in the ionized form. When operated at 120-h detention time of raw wastewater, the high-rate oxidation pond maintained a steady state with respect to algal growth and oxygen concentration, and the concentration of ammonia did not exceed 1.0 mM. Shifting the pond to 48-h detention time caused an increase in ammonia concentration in the pond water to 2.5 mM, and the pond gradually turned anaerobic. Photosynthesis, which usually elevates the pH of the pond water to 9.0 to 10.0, could not proceed beyond pH 7.9 because of the high concentration of ammonia, and the algal population was washed out and reduced to a concentration that could maintain a doubling time of 48 h without photosynthesis bringing the pH to inhibitory levels. Under these conditions, the pH of the bond becomes a factor that limits the operational efficiency of the oxidation pond.     

Effect of pH on Inorganic Carbon Uptake in Algal Cultures

Biomass production by the green algae Scenedesmus obliquus and Chlorella vulgaris in intensive laboratory continuous cultures was considerably affected by the pH at which the cultures were maintained. Carbon photoassimilation experiments revealed that pH values in the range of 8 to 9 were important for determining the free CO₂ concentrations in the medium. With higher pH values, additional pH effects were observed involving a decrease in the relative high affinity of low CO₂-adapted algae to free CO₂. The carbon uptake rate by high CO₂-adapted algae after transfer to low free CO₂ medium was characterized by a lag period of about 30 min, after which the affinity of the algae to CO₂ increased considerably. Both continuous growth and carbon uptake experiments indicated that artificially maintained high free CO₂ concentrations are recommended for maximal production in intensive outdoor algal cultures.     

Seasonal patterns of phytoplankton productivity and abundance in nearshore oligotrophic waters of the Levant Basin (Mediterranean)

A biweekly sampling program from two stations at the easternLevant Basin was carried out during a 1-year period (1983).The first station (neritic) was located 2 km offshore over theIsraeli continental shelf, while the second (pelagic) was located10 km offshore slightly beyond the continental shelf. It wasfound that during summer the relatively close pelagic watershad chlorophyll a concentrations comparable with the most oligotrophicdeep sea regions of the world's oceans. During winter and spring,profound fluctuations were observed in both phytoplankton standingcrop and primary productivity at the neritic station. This wasin response to weather phenomena, such as heavy rains or storms,which did not affect the pelagic Station to such an extent.The picoplanklon size fraction (<3 µm) dominated atthe neritic station during summer and fall, while the nanoplanktonfraction (3–20 µm) dominated during spring. At thepelagic station the picoplankton fraction dominated almost allyear round, but it is suspected that some portion of it wasphotosynthetically inactive.     

Mechanisms of the mediated neutrophil reactivity to Escherichia coli peptidoglycan

The opsonic properties of normal serum with respect to E. coli peptidoglycan was studied under the actual conditions of the oxygen-dependent metabolism of neutrophils. In the course of the differentiated study of the influence of antibodies, the classical and the alternative cascades of complement the serum was heated, treated with ethylenediaminetetraacetate and ethylene glycol tetraacetate, exhausted in the cold. In serial experiments the opsonic activity of purified fibronectin was studied. The indirect reactions were shown to be the leading mechanisms of the neutrophil-stimulated activity of E. coli peptidoglycan. IgG was found to be in the center of the opsonic cooperation and thus to determine the quantitative manifestation of the total phenomenon. Complement proved to be of lesser importance; depending on the conditions of the experiment, the activation of complement occurred by the alternative way (after the removal of antibodies) or the classical way (whole serum). The actual contribution of IgG-independent and complement-independent opsonins was insignificant. Fibronectin in physiological concentrations showed no opsonic activity.     

Reactivity to bacterial peptidoglycans in a phagocytosis system. 1. The range of immunological specificity of Staphylococcus aureus peptidoglycan

The antigenic features of S. aureus peptidoglycan (PG) were studied in the reaction of stimulation of oxygen-dependent neutrophil metabolism, mediated by the IgG opsonins of normal human serum. The study was carried out at different taxonomic levels: the species (S. aureus), the genus (Staphylococcus), the family (Micrococcaceae), as well as in relation to remote taxons (organisms belonging to the families Streptococcaceae, Enterobacteriaceae, Neisseriaceae, to the genus Corynebacterium). All S. aureus strains were identical with respect to the specificity of their PG, essentially differing from other bacteria in this regard. After the removal of antibodies to different PG the effectiveness of PG opsonization decreased by 10.4-44.7%. Such decrease was most pronounced in experiments with the PG of streptococci (S. pyogenes, S. faecalis, S. salivarius) and Micrococcus luteus.     

Effect of Hard Detergents on Algae in a High-Rate-Oxidation Pond

Regular concentrations of hard detergents in domestic wastewater do not affect algal growth in a high-rate-oxidation pond. The addition of nonionic hard detergent at concentrations above 60 mg/liter decreased the algal concentration in the batch culture, and complete lysis of algal cells was observed within a few days at a detergent concentration of 100 mg/liter.     

Toxicity of ammonia to algae in sewage oxidation ponds.

Ammonia, at concentrations over 2.0 mM and at pH values over 8.0, inhibits photosynthesis and growth of Scenedesmus obliquus, a dominant species in high-rate sewage oxidation ponds. Photosynthesis of Chlorella pyrenoidosa, Anacystis nidulans, and Plectonema boryanum is also susceptible to ammonia inhibition. Dark respiration and cell morphology were unaffected by any combination of pH and ammonia concentrations tested, thus limiting the apparent effect to inhibition of the normal function of the chloroplasts. Methylamine had the same effect as ammonia, and its penetration into the cells was found to be pH dependent. Therefore, the dependence of toxicity of amines to algae on pH apparently results from the inability to penetrate the cell membrane in the ionized form. When operated at 120-h detention time of raw wastewater, the high-rate oxidation pond maintained a steady state with respect to algal growth and oxygen concentration, and the concentration of ammonia did not exceed 1.0 mM. Shifting the pond to 48-h detention time caused an increase in ammonia concentration in the pond water to 2.5 mM, and the pond gradually turned anaerobic. Photosynthesis, which usually elevates the pH of the pond water to 9.0 to 10.0, could not proceed beyond pH 7.9 because of the high concentration of ammonia, and the algal population was washed out and reduced to a concentration that could maintain a doubling time of 48 h without photosynthesis bringing the pH to inhibitory levels. Under these conditions, the pH of the bond becomes a factor that limits the operational efficiency of the oxidation pond.     

Algae/Bacteria Ratio in High-Rate Ponds Used for Waste Treatment

Algae, bacteria, and zooplankton were counted in samples drawn from 120- and 150-m² high-rate algae ponds (those used for wastewater treatment). The fraction of nondegraded organic matter was estimated by comparing the ratio of biological and chemical oxygen demands and the bacterial, algal, and zooplankton counts to volatile suspended solids. With pond effluent quality at an acceptable level (around 18 mg of dissolved biological oxygen demand), the algae/bacteria ratio was around 1:100 or even higher, the zooplankton count was negligible, and the bacterial concentration was approximately 10¹¹ cells per liter by direct count. The data for bacteria exceeded those of earlier studies by one to three orders of magnitude.