Energy from vascular plant wastewater treatment systems

Water hyacinth (Eichhornia crassipes), duckweed (Spirodela sp. andLemna sp.), water pennywort (Hydrocotyle ranunculoides), and kudzu (Pueraria lobata) were anaerobically fermented using an anaerobic filter technique that reduced the total digestion time from 90 d to an average of 23 d and produced 0.14-0.22 m³ CH₄/kg (dry weight) (2.3-3.6 ft³/lb) from mature filters for the 3 aquatic species. Kudzu required an average digestion time of 33 d and produced an average of 0.21 m³ CH₄/kg (dry weight) (3.4 ft³/lb). The anaerobic filter provided a large surface area for the anaerobic bacteria to establish and maintain an optimal balance of facultative, acid-forming, and methane-producing bacteria. Consequently the efficiency of the process was greatly improved over prior batch fermentations.     

Growing duckweed in swine wastewater for nutrient recovery and biomass production

Spirodela oligorrhiza, a promising duckweed identified in previous studies, was examined under different cropping conditions for nutrient recovery from swine wastewater and biomass production. To prevent algae bloom during the start-up of a duckweed system, inoculating 60% of the water surface with duckweed fronds was required. In the growing season, the duckweed system was capable of removing 83.7% and 89.4% of total nitrogen (TN) and total phosphorus (TP) respectively from 6% swine lagoon water in eight weeks at a harvest frequency of twice a week. The total biomass harvested was 5.30 times that of the starting amount. In winter, nutrients could still be substantially removed in spite of the limited duckweed growth, which was probably attributed to the improved protein accumulation of duckweed plants and the nutrient uptake by the attached biofilm (algae and bacteria) on duckweed and walls of the system.     

Duckweed (Lemna minor) as a model plant system for the study of human microbial pathogenesis

Background

Plant infection models provide certain advantages over animal models in the study of pathogenesis. However, current plant models face some limitations, e.g., plant and pathogen cannot co-culture in a contained environment. Development of such a plant model is needed to better illustrate host-pathogen interactions.

Methodology/Principal Findings

We describe a novel model plant system for the study of human pathogenic bacterial infection on a large scale. This system was initiated by co-cultivation of axenic duckweed (Lemna minor) plants with pathogenic bacteria in 24-well polystyrene cell culture plate. Pathogenesis of bacteria to duckweed was demonstrated with Pseudomonas aeruginosa and Staphylococcus aureus as two model pathogens. P. aeruginosa PAO1 caused severe detriment to duckweed as judged from inhibition to frond multiplication and chlorophyll formation. Using a GFP-marked PAO1 strain, we demonstrated that bacteria colonized on both fronds and roots and formed biofilms. Virulence of PAO1 to duckweed was attenuated in its quorum sensing (QS) mutants and in recombinant strains overexpressing the QS quenching enzymes. RN4220, a virulent strain of S. aureus, caused severe toxicity to duckweed while an avirulent strain showed little effect. Using this system for antimicrobial chemical selection, green tea polyphenols exhibited inhibitory activity against S. aureus virulence. This system was further confirmed to be effective as a pathogenesis model using a number of pathogenic bacterial species.

Conclusions/Significance

Our results demonstrate that duckweed can be used as a fast, inexpensive and reproducible model plant system for the study of host-pathogen interactions, could serve as an alternative choice for the study of some virulence factors, and could also potentially be used in large-scale screening for the discovery of antimicrobial chemicals.     

Effects of nitrate treatment on a mixed species, oil field microbial biofilm

Biofilms of bacteria, indigenous to oil field produced water, were grown in square section, glass capillary flow cells at 45 °C. Initially, in situ image analysis microscopy revealed predominantly coccoid bacteria (length-to-width ratio measurements (l _c:w _c) of bacterial cells gave a mean value of 1.1), while chemical measurements confirmed sulphate reduction and sulphide production. After nitrate ion addition at 100 and 80 mg/l, in the two repeat experiments respectively, the dominance of rod-shaped bacteria (mean l _c:w _c = 2.8) was observed. This coincided with the occurrence of nitrate reduction in the treated flow cells. Beneficially, no significant increase in biofilm cover was observed after the addition of nitrate. The dominant culturable nitrate-reducing bacterium was Marinobacter aquaeolei. The l _c:w _c ratio measured here concurs with previously reported cell dimensions for this organism. Several Marinobacter strains were also isolated from different oil fields in the North Sea where nitrate treatment has been applied to successfully treat reservoir souring, implying that this genus may play an important role in nitrate treatment.     

Nitrogen Fixation Dynamics of Two Diazotrophic Communities in Mono Lake, California

Two types of diazotrophic microbial communities were found in the littoral zone of alkaline hypersaline Mono Lake, California. One consisted of anaerobic bacteria inhabiting the flocculent surface layers of sediments. Nitrogen fixation (acetylene reduction) by flocculent surface layers occurred under anaerobic conditions, was not stimulated by light or by additions of organic substrates, and was inhibited by O₂, nitrate, and ammonia. The second community consisted of a ball-shaped association of a filamentous chlorophyte (Ctenocladus circinnatus) with diazotrophic, nonheterocystous cyanobacteria, as well as anaerobic bacteria (Ctenocladus balls). Nitrogen fixation by Ctenocladus balls was usually, but not always, stimulated by light. Rates of anaerobic dark fixation equaled those in the light under air. Fixation in the light was stimulated by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea and by propanil [N-(3,4-dichlorophenyl)propanamide]. 3-(3,4-Dichlorophenyl)-1,1-dimethyl urea-elicited nitrogenase activity was inhibited by ammonia (96%) and nitrate (65%). Fixation was greatest when Ctenocladus balls were incubated anaerobically in the light with sulfide. Dark anaerobic fixation was not stimulated by organic substrates in short-term (4-h) incubations, but was in long-term (67-h) ones. Areal estimates of benthic N₂ fixation were measured seasonally, using chambers. Highest rates (~29.3 μmol of C₂H₄ m⁻² h⁻¹) occurred under normal diel regimens of light and dark. These estimates indicate that benthic N₂ fixation has the potential to be a significant nitrogen source in Mono Lake.     

Common Duckweed (Lemna minor) Is a Versatile High-Throughput Infection Model For the Burkholderia cepacia Complex and Other Pathogenic Bacteria

Members of the Burkholderia cepacia complex (Bcc) have emerged in recent decades as problematic pulmonary pathogens of cystic fibrosis (CF) patients, with severe infections progressing to acute necrotizing pneumonia and sepsis. This study presents evidence that Lemna minor (Common duckweed) is useful as a plant model for the Bcc infectious process, and has potential as a model system for bacterial pathogenesis in general. To investigate the relationship between Bcc virulence in duckweed and Galleria mellonella (Greater wax moth) larvae, a previously established Bcc infection model, a duckweed survival assay was developed and used to determine LD₅₀ values. A strong correlation (R² = 0.81) was found between the strains’ virulence ranks in the two infection models, suggesting conserved pathways in these vastly different hosts. To broaden the application of the duckweed model, enteropathogenic Escherichia coli (EPEC) and five isogenic mutants with previously established LD₅₀ values in the larval model were tested against duckweed, and a strong correlation (R² = 0.93) was found between their raw LD₅₀ values. Potential virulence factors in B. cenocepacia K56-2 were identified using a high-throughput screen against single duckweed plants. In addition to the previously characterized antifungal compound (AFC) cluster genes, several uncharacterized genes were discovered including a novel lysR regulator, a histidine biosynthesis gene hisG, and a gene located near the gene encoding the recently characterized virulence factor SuhB_Bc. Finally, to demonstrate the utility of this model in therapeutic applications, duckweed was rescued from Bcc infection by treating with bacteriophage at 6-h intervals. It was observed that phage application became ineffective at a timepoint that coincided with a sharp increase in bacterial invasion of plant tissue. These results indicate that common duckweed can serve as an effective infection model for the investigation of bacterial virulence factors and therapeutic strategies to combat them.     

Survivorship of juvenile surf clams Donax serra (Bivalvia, Donacidae) exposed to severe hypoxia and hydrogen sulphide

Toxic “sulphide eruptions” sporadically occur in the highly productive inshore regions of the central Namibian Benguela upwelling system. The surf clam Donax serra (Röding, 1798) dominates the intertidal and upper subtidal of large exposed sandy beaches of southern Africa and its recruitment seems to be affected by sulphide events. The reaction of juvenile surf clams to low oxygen concentrations and sulphide occurrence (0.1 mmol l⁻¹) was examined by in vitro exposure experiments in a gas-tight continuous flow system. After 2 h of hypoxic- and hypoxic/sulphidic conditions, clams moved to the sediment surface, aiding their passive transport to areas with more favourable conditions. The clams showed a high sulphide detoxification capacity by oxidising the penetrating hydrogen sulphide to non-toxic thiosulphate. Moreover, juvenile D. serra switched to anaerobic energy production, indicated by the significant accumulation of succinate and, to some extent, alanine. Test animals were not able to reduce their energy requirements enough to withstand long periods of exposure, leading to a median survival time (LT₅₀) of 80 h under hypoxic sulphide incubation. In conclusion, natural “sulphide eruptions”, especially those with a large spatial and temporal extension, have to be considered as an important factor for D. serra recruitment failures. Hydrogen sulphide is assumed to be a potential community structuring factor.     

The effects of the oligochaete Branchiura sowerbyi Beddard (Tubificidae) on the biological and chemical characteristics of overlying water and soil in a submerged ricefield soil system

Submerged ricefield soils, outdoors, and with and without the tubificid Branchiura sowerbyi Beddard, were used to study the effects of tubificids on some biotic and abiotic variables. Measurements of chemical and biological features showed that the tubificids accelerated the diffusion of dissolved substances, increased the release of soil bacteria and enhanced the production of algae and a duckweed (Lemna paucicostata) in the overlying water. In the presence of tubificids, increased numbers of Cladocera (Moina sp., Simocephalus sp.) and ostracods in the overlying water, and higher number of sulfate-reducing bacteria in the soil were observed. The tubificids further enhanced the microbial activity in the upper soil layer, which in turn produced large amounts of organic matter.     

Effect of anaerobiosis on photosynthetic reactions and nitrogen metabolism of algae with and without hydrogenase

Summary The effect of anaerobic (N₂+CO₂) pre-incubation in the dark on photosynthetic reactions (O₂ evolution, measured manometrically and with the oxygraph; fluorescence; and photoproduction of H₂, measured with the mass spectrometer) was studied in algae with hydrogenase (strains of Chlorella fusca, C. kessleri, C. vulgaris f. tertia, and Ankistrodesmus braunii) and in algae without hydrogenase (strains of Chlorella vulgaris, C. saccharophila, and C. minutissima).The inhibition by anaerobic incubation of photosynthetic O₂ evolution is much stronger in algae without hydrogenase than it is in algae with hydrogenase. The effect of anaerobiosis is most pronounced at rather low light intensity (about 1000 lux), in acid medium (pH 4), and after prolonged anaerobic incubation in the dark (about 20 h). These results indicate that the presence of hydrogenase might be ecologically advantageous for algae under certain conditions.Chlorophyll fluorescence showed the fastest response to anaerobic incubation, and the most pronounced difference between algae with and without hydrogenase. After only 30 min under N₂+CO₂, fluorescence in algae with hydrogenase starts with a peak and decreases within 10 to 20 sec to a rather low steady-state level which is only slightly higher than that found under aerobic conditions. In algae without hydrogenase, fluorescence is rather low during the first 1 to 2 sec and then rises to a higher steady-state level which is much higher than that of the aerobic controls. This indicates an inhibition due to anaerobiosis of photosystem II in algae without hydrogenase.Algae with hydrogenase can react in different ways during the first minutes of illumination. In some cases there is an immediate photoproduction of H₂, which is followed after a few minutes by photosynthetic O₂ evolution; in other algae there is a simultaneous production of H₂ and O₂ from the very beginning; in a few experiments there was no photoproduction of H₂ at all, and in this case there was no photosynthetic O₂ evolution either. Thus, photoproduction of H₂ seems to be the process which normally enables algae with hydrogenase to oxidise and thereby activate their photosynthetic electron transport system after anaerobic incubation.A mass spectrometric search for nitrogen fixation (using N₂ and acetylene) in eucaryotic green algae gave negative results, even with species containing hydrogenase and under anaerobic conditions.     

Decomposition of Blue-Green Algal (Cyanobacterial) Blooms in Lake Mendota, Wisconsin

Decomposition of natural populations of Lake Mendota phytoplankton dominated by blue-green algae (cyanobacteria) was monitored by using oxygen uptake and disappearance of chlorophyll, algal volume (fluorescence microscopy), particulate protein, particulate organic carbon, and photosynthetic ability (¹⁴CO₂ up-take). In some experiments, decomposition of ¹⁴C-labeled axenic cultures of Anabaena sp. was also measured. In addition to decomposition, mineralization of inorganic nitrogen and phosphorus were followed in some experiments. Decomposition could be described as a first-order process, and the rate of decomposition was similar to that found by others using pure cultures of eucaryotic algae. Nitrogen and phosphorus never limited the decomposition process, even when the lake water was severely limited in soluble forms of these nutrients. This suggests that the bacteria responsible for decomposition can obtain all of their key nutrients for growth from the blue-green algal cells. Filtration of lake water through plankton netting that removed up to 90% of the algal biomass usually did not cause a similar decrease in oxygen demand, suggesting that most of the particulate organic matter used for respiration of the decomposing bacteria was in a small-particle fraction. Short-term oxygen demand correlated well with the particulate chlorophyll concentration of the sample, and a relationship was derived that could be used to predict community respiration of the lake from chlorophyll concentration. Kinetic analysis showed that not all analyzed components disappeared at the same rate during the decomposition process. The relative rates of decrease of the measured parameters were as follows: photosynthetic ability > algal volume > particulate chlorophyll > particulate protein. Decomposition of ¹⁴C-labeled Anabaena occurred at similar rates with aerobic epilimnetic water and with anaerobic sediment, but was considerably slower with anaerobic hypolimnetic water. Of the various genera present in the lake, Aphanizomenon and Anabaena were more sensitive to decomposition than was Microcystis. In addition to providing a general picture of the decomposition process, the present work relates to other work on sedimentation to provide a detailed picture of the fate of blue-green algal biomass in a eutrophic lake ecosystem.     

Bacterial communities of some brown and red algae from Peter the Great Bay, the Sea of Japan

The structure of microbial communities of brown algae, red algae, and of the red alga Gracilaria verrucosa, healthy and affected with thallus rot, were comparatively investigated; 61 strains of heterotrophic bacteria were isolated and characterized. Most of them were identified to the genus level, some Vibrio spp., to the species level according to their phenotypic properties and the fatty acid composition of cellular lipids. The composition of the microflora of two species of brown algae was different. In Chordaria flagelliphormis, Pseudomonas spp. prevailed, and in Desmarestia viridis, Bacillus spp. The composition of the microflora of two red algae, G. verrucosa and Camphylaephora hyphaeoides, differed mainly in the ratio of prevailing groups of bacteria. The most abundant were bacteria of the CFB cluster and pseudoalteromonads. In addition, the following bacteria were found on the surface of the algae: Sulfitobacter spp., Halomonas spp., Acinetobacter sp., Planococcus sp., Arthrobacter sp., and Agromyces sp. From tissues of the affected G. verrucosa, only vibrios were isolated, both agarolytic and nonagarolytic. The existence of specific bacterial communities characteristic of different species of algae is suggested and the relation of Vibrio sp. to the pathological process in the tissues of G. verrucosa is supposed.     

Effects of available surface on gaseous emissions from group-housed gestating sows kept on deep litter

In the European Union, the group-housed pregnant sows have to have a minimal legal available area of 2.25 m²/sow. However, it has been observed that an increased space allowance reduces agonistic behaviour and consecutive wounds and thus induces better welfare conditions. But, what about the environmental impacts of this greater available area? Therefore, the aim of this study was to quantify pollutant gases emissions (nitrous oxide, N₂O, methane, CH₄, carbon dioxide, CO₂ and ammonia, NH₃), according to the space allowance in the raising of gestating sows group-housed on a straw-based deep litter. Four successive batches of 10 gestating sows were each divided into two homogeneous groups and randomly allocated to a treatment: 2.5 v. 3.0 m²/sow. The groups were separately kept in two identical rooms. A restricted conventional cereals based diet was provided once a day in individual feeding stalls available only during the feeding time. Rooms were automatically ventilated. The gas emissions were measured by infra red photoacoustic detection during six consecutive days at the 6th, 9th and 12th weeks of gestation. Sows performance (body weight gain, backfat thickness, number and weight of piglets) was not significantly different according to the space allowance. In the room with 3.0 m²/sow and compared with the room with 2.5 m²/sow, gaseous emissions were significantly greater for NH₃ (6.29 v. 5.37 g NH₃-N/day per sow; P < 0.01) and significantly lower for N₂O (1.78 v. 2.48 g N₂O-N/day per sow; P < 0.01), CH₄ (10.15 v. 15.21 g/day per sow; P < 0.001), CO₂ equivalents (1.11 v. 1.55 kg/day per sow; P < 0.001), CO₂ (2.12 v. 2.41 kg/day per sow; P < 0.001) and H₂O (3.10 v. 3.68 kg/day per sow; P < 0.001). In conclusion, an increase of the available area for group-housed gestating sow kept on straw-based deep litter seems to be ambiguous on an environmental impacts point of view. Compared with a conventional and legal available area, it favoured NH₃ emissions, probably due to an increased emitting surface. However, about greenhouse gases, it decreased N₂O, CH₄ and CO₂ emissions, probably due to reduced anaerobic conditions required for their synthesis, and led to a reduction of CO₂ equivalents emissions.     

Dynamics of Phototrophic Microbial Populations in the Chemocline of a Meromictic Basin of Lake Banyoles

The ecological succession of two microbial planktonic populations in the chemocline of C-IV, a meromictic basin of Lake Banyoles has been studied during the stratification period of 1989 (June to October). A dense population of deep-living algae Cryptomonas phaseolus was found growing between 13 and 14 meters depth, forming peaks of 10 to 20 μg.l⁻¹ of chlorophyll a. The physical and chemical properties of the water were continually changing during the studied period and were strongly related to the biological processes developed at the monimolimnion of C-IV. Although the monimolimnion of the basin was initially anoxic (Eh values were around +300 mV and sulphide was not present at detectable levels) soluble iron and sulphide appeared sequentially, displacing the deep-living algae population and setting the conditions for the development of a population of brown Chlorobiaceae, Chlorobium phaeobacteroides. Although Cryptomonas population abandons the chemocline (13.75 m.) and decreases its biomass, it does not disappear at all, but moves upwards (8–11 m.) leaving the zone to sulphur phototrophic bacteria which find optimal conditions to grow. The sulphide diffusion coming from sediment and the nutrient limitation were the main responsible factors accounting for the upwards migration and the decrease of growth rate of the algal population respectively.     

Formation of N-Carboxymethyl Amino Acid from the Reaction of α-Amino Acid with Glyoxal

Enrichment cultures in a medium containing 0.1% methanol and 0.1% bicarbonate at pH 7.0 under anaerobic conditions in the light became mainly green in color. Forty-four enrichment cultures, which showed abundant growth, were obtained from 46 different sources and found to contain cells of methanol-utilizing bacteria and green algae as predominant members. From these enrichment cultures, two strains of bacteria and two strains of algae were isolated. The microorganisms isolated were designated as bacterium No. 7, bacterium No. 8, Chlorella sp. A-1 and Chlorella sp. B-1, respectively. Stable mixed cultures were easily formed by mixing the isolated cultures of bacteria and algae. Both methanol and bicarbonate were necessary for the growth of the mixed cultures under anaerobic-light conditions. Growth behavior of the mixed cultures was examined on a medium containing 0.1% methanol and 0.1 % bicarbonate at 30°C in the light (about 6000 lx). The maximum specific growth rate for the cultures, µ_max, was 0.092 hr^?1 (doubling time, 7.5 hr). The maximum cell yield was 0.87 g dry-cell weight per g of methanol used. The protein content of the biomass was 65%.     

Rates of Transformation of Methyl Parathion and Diethyl Phthalate by Aufwuchs Microorganisms

Using batch cultures, we determined transformation rates for low concentrations of two toxicants—an insecticide, methyl parathion (O,O-dimethyl O-p-nitrophenyl phosphorothioate), and a plasticizer, diethyl phthalate—by aufwuchs, aquatic microbial growth attached to submerged surfaces or suspended in streamers or mats. Aufwuchs samples were collected from field sites, an indoor channel, and a continuous-flow fermentor. Aufwuchs fungi, protozoa, and algae did not transform methyl parathion or diethyl phthalate, but bacteria rapidly transformed both chemicals. Second-order transformation rate coefficients, K_b, based on total plate counts of bacteria in aufwuchs, were determined for potential use in a mathematical model capable of predicting the transport and fate of chemicals in aquatic systems. K_b for both methyl parathion and diethyl phthalate decreased as the concentration of total bacteria, [B], increased in aufwuchs. This effect resulted from the proportion of nontransformer to transformer bacteria increasing as [B] increased and from the rate of transformation per transformer cell decreasing as [B] increased. First-order transformation rate coefficients, K₁, were relatively stable per unit of surface area colonized by aufwuchs, because K_b decreased as [B] increased (K₁ = K_b × [B]).     

海水中藻菌共培养体系对碳氮磷的吸收转化

海洋环境中,细菌和微藻之间的物质交换是生源要素在自然界中迁移转化的重要方式。为进一步了解生源要素的生物地球化学循环,在实验室模拟条件下,研究了共培养体系中营养盐和有机物在细菌和微藻之间的转换。通过纯培养中肋骨条藻(Skeletonema costatum)、东海原甲藻(Prorocentrum donghaiense)、天然海水中的细菌以及藻菌混合培养,分析了营养盐和有机物随藻菌生物量的变化情况,并计算了溶解有机碳(DOC)和溶解有机氮(DON)的浓度比值[(DOC/DON)a]。结果发现,在共培养体系中,细菌对中肋骨条藻的生长有抑制作用,对东海原甲藻影响不明显;中肋骨条藻有利于细菌生长,东海原甲藻抑制细菌生长,这种不同可能与微藻的粒径有关。海洋细菌在2种藻的指数生长均期均会促进微藻吸收氨氮(NH_4-N),但在生长末期NH_4-N以释放为主。硝氮(NO_3-N)的浓度与藻的生长呈负相关,但在衰亡期NO_3-N略有增加,表明NO_3-N再生所需时间较长。细菌对硝氮的吸收量较少,但对其再生有贡献。细菌和中肋骨条藻对磷酸盐(PO_4-P)的吸收存在竞争,但与东海原甲藻的竞争关系不明显。不同培养体系中DOC浓度变化不同,在藻菌共培养体系中增加较快,纯藻培养体系中增加缓慢,在纯菌培养体系中缓慢减少。通过对DOC与DON浓度比值的分析,发现用判断颗粒有机碳(POC)来源的方法可以分析DOC的来源。     

Start-up performance and granular sludge features of an improved external circulating anaerobic reactor for algae-laden water treatment

The microbial characteristics of granular sludge during the rapid start of an enhanced external circulating anaerobic reactor were studied to improve algae-laden water treatment efficiency. Results showed that algae laden water was effectively removed after about 35 d, and the removal rates of chemical oxygen demand (COD) and algal toxin were around 85% and 92%, respectively. Simultaneously, the gas generation rate was around 380 mL/gCOD. The microbial community structure in the granular sludge of the reactor was complicated, and dominated by coccus and filamentous bacteria. Methanosphaera, Methanolinea, Thermogymnomonas, Methanoregula, Methanomethylovorans, and Methanosaeta were the major microorganisms in the granular sludge. The activities of protease and coenzyme F₄₂₀ were high in the granular sludge. The intermittent stirring device and the reverse-flow system were further found to overcome the disadvantage of the floating and crusting of cyanobacteria inside the reactor. Meanwhile, the effect of mass transfer inside the reactor can be accelerated to help give the reactor a rapid start.     

Nutrient availability affects the response of the calcifying chlorophyte Halimeda opuntia (L.) J.V. Lamouroux to low pH

Atmospheric carbon dioxide emissions cause a decrease in the pH and aragonite saturation state of surface ocean water. As a result, calcifying organisms are expected to suffer under future ocean conditions, but their physiological responses may depend on their nutrient status. Because many coral reefs experience high inorganic nutrient loads or seasonal changes in nutrient availability, reef organisms in localized areas will have to cope with elevated carbon dioxide and changes in inorganic nutrients. Halimeda opuntia is a dominant calcifying primary producer on coral reefs that contributes to coral reef accretion. Therefore, we investigated the carbon and nutrient balance of H. opuntia exposed to elevated carbon dioxide and inorganic nutrients. We measured tissue nitrogen, phosphorus and carbon content as well as the activity of enzymes involved in inorganic carbon uptake and nitrogen assimilation (external carbonic anhydrase and nitrate reductase, respectively). Inorganic carbon content was lower in algae exposed to high CO₂, but calcification rates were not significantly affected by CO₂ or inorganic nutrients. Organic carbon was positively correlated to external carbonic anhydrase activity, while inorganic carbon showed the opposite correlation. Carbon dioxide had a significant effect on tissue nitrogen and organic carbon content, while inorganic nutrients affected tissue phosphorus and N:P ratios. Nitrate reductase activity was highest in algae grown under elevated CO₂ and inorganic nutrient conditions and lowest when phosphate was limiting. In general, we found that enzymatic responses were strongly influenced by nutrient availability, indicating its important role in dictating the local responses of the calcifying primary producer H. opuntia to ocean acidification.     

Microbial populations regulate greenhouse gas emissions in Sundarban mangrove ecosystem,India

Mangrove ecosystems are significant sources of greenhouse gases (GHG) that is attributed to microbial activity. However, it is still unknown how the sediment microbial populations affect GHG emissions in mangrove ecosystem. Since little is known about microbial populations of mangroves, the present study was aimed to understand the structure and function of microbial communities in the Indian part of the Sundarban mangrove ecosystem in relation to environmental variables and variation of GHG emissions during three seasons: pre-monsoon (March–June), monsoon (July–October) and post-monsoon (November–February).Seasonal variations of carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O) gas samples were taken from the mangrove bed. Culture methods were used to detect twelve different types of microbes such as heterotrophic (Htp), N₂ fixing (Nfix), nitrifying (Ntfn), sulfur oxidizing (Soxd), Gram-negative (GMn), Gram-positive (GMp), spore forming (Sfor), denitrifying (DNtfn), anaerobic (Anrb), phosphate solubilizing (Psol), cellulose degrading (Cdeg) bacteria and actinomycetes (Actm). In the monsoon, populations of the Htp, Anrb, Psol, and Cdeg bacteria were more prevalent, whereas populations of the GMn, GMp, Ntfn, DNtfn bacteria, and Actm bacteria were more prevalent in the post-monsoon. Monsoonal CO₂ and CH₄ fluxes were larger than pre-monsoon and post-monsoon, resulting in increased microbial soil respiration and breakdown of soil organic carbon. Because of higher denitrification and soil temperature, N₂O flux was higher in the pre-monsoon period, followed by monsoon and post-monsoon periods. A univariate statistical correlation was employed to assess the relationships between environmental variables and different microbial populations. An ANN (artificial neural network) model was proposed to evaluate the relevance of microbial population contribution to GHG emissions, and it indicated that the Htp, Anrb, and Dntfn microbial populations were most relevant for CO₂, CH₄, and N₂O emissions. The suggested model would be used to assess the drivers behind GHG emission in the mangroves located at different parts of the world.