Airborne remote sensing of chlorophyll distributions in Mono Lake, California

As part of a long-term investigation of seasonal and interannual variations of plankton in Mono Lake (California), we developed a methodology using airborne imaging spectrometry to synoptically measure chlorophyll concentrations. Images of Mono Lake were acquired with NASA's Airborne Visible and Infrared Imaging Spectrometer (AVIRIS) and were atmospherically corrected by applying a version of the radiative transfer model MODTRAN. Using a predictive equation for calculation of chlorophyll based on a band ratio of remote sensing reflectances (R _{{ rs}}; R _{{ rs}} 490 nm/R _{{ rs}} 550 nm), spatial distributions of chlorophyll throughout the lake were determined; broad east to west gradients in chlorophyll and gyres are evident.     

Stratification of microbial assemblages in Mono Lake, California, and response to a mixing event

Vertical profiles of microbial assemblages from samples of Mono Lake water collected in July 1994 and in April and July 1995 were obtained by analyzing DNA via the polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) of 16S ribosomal RNA (rRNA) genes. The microbial assemblage was vertically stratified and distributions of individual ribotypes were coherent with temperature, salinity, irradiance and dissolved oxygen distributions at the beginning of the study in July of 1994. The lake mixed completely during the winter of 1994–1995 and was beginning to stratify thermally by April 1995. Water column gradients were weak and oxygen was depleted at depth. The microbial assemblage was uniformly distributed throughout the water column except at 20 m, where one band dominated. The microbial assemblage was vertically stratified again by July 1995. Partial sequences (134–160 bp, except one of 83 bp) obtained from DGGE bands revealed affinities to known organisms, but only one potentially exact match was found. With a few exceptions, the same ribotypes were present on all sampling dates; there was no evidence for a marked seasonal succession in microbial community composition, despite the dramatic changes in limnological conditions that accompanied the winter overturn. A band that was ubiquitous in samples from the oxycline and hypolimnion in July of both years was found throughout the water column in April. This sequence could be attributed to the chloroplast rRNA gene of an unusual phytoplankter, the green alga Picocystis salinarum.     

Chlorophyll differences in Mono Lake (California) observable on Landsat imagery

In Mono Lake (California), a large saline lake, chlorophyll concentrations in the euphotic zone increased from 4 to 45 µg l^–1 between July and October 1979. These seasonal changes in chlorophyll are detectable on imagery obtained with the multispectral scanner on Landsat. Computer-compatible tapes of Landsat images were normalized for solar zenith and corrected for atmospheric scatter and absorption to obtain Landsat band 4 emittances (W m^–2 str^–1) of 13.4 ± 0.5 when chlorophyll was 4 µg l^–1 and 4.6 ± 0.3 when chlorophyll was 45 µg l^–1. Lake wide, spatial heterogeneity of chlorophyll of 2 µg l^–1 in July and 8 µg l^–1 in October was not detectable on the Landsat imagery.     

Enrichment and isolation of Bacillus beveridgei sp. nov., a facultative anaerobic haloalkaliphile from Mono Lake, California, that respires oxyanions of tellurium, selenium, and arsenic

Mono Lake sediment slurries incubated with lactate and tellurite [Te(IV)] turned progressively black with time because of the precipitation of elemental tellurium [Te(0)]. An enrichment culture was established from these slurries that demonstrated Te(IV)-dependent growth. The enrichment was purified by picking isolated black colonies from lactate/Te(IV) agar plates, followed by repeated streaking and picking. The isolate, strain MLTeJB, grew in aqueous Te(IV)-medium if provided with a small amount of sterile solid phase material (e.g., agar plug; glass beads). Strain MLTeJB grew at high concentrations of Te(IV) (~8 mM) by oxidizing lactate to acetate plus formate, while reducing Te(IV) to Te(0). Other electron acceptors that were found to sustain growth were tellurate, selenate, selenite, arsenate, nitrate, nitrite, fumarate and oxygen. Notably, growth on arsenate, nitrate, nitrite and fumarate did not result in the accumulation of formate, implying that in these cases lactate was oxidized to acetate plus CO₂. Strain MLTeJB is a low G + C Gram positive motile rod with pH, sodium, and temperature growth optima at 8.5–9.0, 0.5–1.5 M, and 40°C, respectively. The epithet Bacillus beveridgei strain MLTeJB^T is proposed.     

Osmoregulation in nestling California gulls at Mono Lake,California

• 1.1. Hematocrit (Hct), plasma sodium ([Na]_pl), chloride ([Cl]_pl) and osmotic concentration (Osm_pl); volume and concentration of 1.0MNaCl induced salt gland secretion (SGS); and weights of osmoregulatory organs: kidneys, adrenal glands, and salt glands and nonosmoregulatory organs (liver and heart) were determined in nestling California gulls, Larus californiens (CG), on Krakatoa Islet, Mono Lake, California.
• 2.2. The mean Hct was 40.0% + 1.0%, the mean [Na]_pl and [Cl]_pl were 153.9 ± 0.9 and 110.1 ± 0.5 mM (n = 22); and the mean Osm_pl was 323.6 ± 1.3 mOsm/kg (n = 18).
• 3.3. In CG nestlings with a mean age of 10 days (n = 7), the mean SGS [Na] was 719 ± 19 mM and the birds secreted 81 ± 18% of the injected fluid containing 59 ± 13% of the injected Na. By the mean age of 23 days (n = 7), mean SGS [Na] was slightly higher (790 ± 30 mM than in younger birds (P < 0.05), but the percentage of secreted fluid (54 ± 10%) and Na (42 ± 6%) tended to be less.
• 4.4. In 18 CG nestlings mean organ weights (% body weight) were: kidneys 1.52 ±0.06%; salt glands, 0.13 ±0.01%, and adrenal glands 0.07 ±0.01%.
• 5.5. Nestling CG had significantly greater Hct (P < 0.001), [Na]_pl and [ci]_pl (P < 0.001), Osm_pl, (P < 0.005), and adrenal gland weight (P < 0.01), compared to nestling glaucous-winged Gulls (GWG), L. glaucescens (Hughes, 1984), which nest under cooler, moister conditions. CG kidney weight was smaller (P < 0.001); salt gland weight and salt excretion were the same as GWG.

     

Depth distribution of microbial diversity in Mono Lake,a meromictic soda lake in California

We analyzed the variation with depth in the composition of members of the domain Bacteria in samples from alkaline, hypersaline, and currently meromictic Mono Lake in California. DNA samples were collected from the mixolimnion (2 m), the base of the oxycline (17.5 m), the upper chemocline (23 m), and the monimolimnion (35 m). Composition was assessed by sequencing randomly selected cloned fragments of 16S rRNA genes retrieved from the DNA samples. Most of the 212 sequences retrieved from the samples fell into five major lineages of the domain BACTERIA: alpha- and gamma-Proteobacteria (6 and 10%, respectively), Cytophaga-Flexibacter-Bacteroides (19%), high-G+C-content gram-positive organisms (Actinobacteria; 25%), and low-G+C-content gram-positive organisms (Bacillus and Clostridium; 19%). Twelve percent were identified as chloroplasts. The remaining 9% represented beta- and delta-Proteobacteria, Verrucomicrobiales, and candidate divisions. Mixolimnion and oxycline samples had low microbial diversity, with only 9 and 12 distinct phylotypes, respectively, whereas chemocline and monimolimnion samples were more diverse, containing 27 and 25 phylotypes, respectively. The compositions of microbial assemblages from the mixolimnion and oxycline were not significantly different from each other (P = 0.314 and 0.877), but they were significantly different from those of chemocline and monimolimnion assemblages (P < 0.001), and the compositions of chemocline and monimolimnion assemblages were not significantly different from each other (P = 0.006 and 0.124). The populations of sequences retrieved from the mixolimnion and oxycline samples were dominated by sequences related to high-G+C-content gram-positive bacteria (49 and 63%, respectively) distributed in only three distinct phylotypes, while the population of sequences retrieved from the monimolimnion sample was dominated (52%) by sequences related to low-G+C-content gram-positive bacteria distributed in 12 distinct phylotypes. Twelve and 28% of the sequences retrieved from the chemocline sample were also found in the mixolimnion and monimolimnion samples, respectively. None of the sequences retrieved from the monimolimnion sample were found in the mixolimnion or oxycline samples. Elevated diversity in anoxic bottom water samples relative to oxic surface water samples suggests a greater opportunity for niche differentiation in bottom versus surface waters of this lake.     

Arsenic in contaminated waters: Biogeochemical cycle, microbial metabolism and biotreatment processes

Arsenic is responsible for the contamination of water supplies in various parts of the world and poses a major risk to human health. Its toxicity and bioavailability depend on its speciation, which in turn, depends on microbial transformations, including reduction, oxidation and methylation. This review describes the development of bioprocesses for the treatment of arsenic-contaminated waters based on bacterial metabolism and biogeochemical cycling of arsenic.     

Arsenic respiration and detoxification by purple non-sulphur bacteria under anaerobic conditions

Two arsenic-resistant purple non-sulphur bacteria (PNSB), Q3B and Q3C, were isolated (from industrial contaminated site and paddy fields) and identified by SSU rRNA gene sequencing as Rhodospirillum and Rhodospirillaceae species, respectively. Maximum arsenic reduction by these PNSB was observed in anaerobic conditions. Rhodospirillum sp. Q3B showed 74.92% (v/v) arsenic reduction while Rhodospirillaceae sp. Q3C reduced arsenic up to 76.67% (v/v) in anaerobic conditions. Rhodospirillaceae sp. Q3C was found to contain highest carotenoid content up to 5.6 mg·g⁻¹. Under anaerobic conditions, the isolates were able to respire arsenic in the presence of lactate, citrate, and oxalate. Rhodospirillum sp. Q3B and Rhodospirillaceae sp. Q3C were also found to produce hydrogen gas. Such diverse bacteria can be useful tools for bioremediation purposes. These bacteria can be further exploited and optimized to treat wastewater containing arsenic along with bio-hydrogen production.     

Analysis of fae and fhcD Genes in Mono Lake, California

Genes for two enzymes of the tetrahydromethanopterin-linked C₁ transfer pathway (fae and fhcD) were detected in hypersaline, hyperalkaline Mono Lake (California), via PCR amplification and analysis. Low diversity for fae and fhcD was noted, in contrast to the diversity previously detected in a freshwater lake, Lake Washington (Washington).     

Analysis of fae and fhcD genes in Mono Lake, California

Genes for two enzymes of the tetrahydromethanopterin-linked C(1) transfer pathway (fae and fhcD) were detected in hypersaline, hyperalkaline Mono Lake (California), via PCR amplification and analysis. Low diversity for fae and fhcD was noted, in contrast to the diversity previously detected in a freshwater lake, Lake Washington (Washington).     

Depth Distribution of Microbial Diversity in Mono Lake, a Meromictic Soda Lake in California

We analyzed the variation with depth in the composition of members of the domain Bacteria in samples from alkaline, hypersaline, and currently meromictic Mono Lake in California. DNA samples were collected from the mixolimnion (2 m), the base of the oxycline (17.5 m), the upper chemocline (23 m), and the monimolimnion (35 m). Composition was assessed by sequencing randomly selected cloned fragments of 16S rRNA genes retrieved from the DNA samples. Most of the 212 sequences retrieved from the samples fell into five major lineages of the domain Bacteria: α- and γ-Proteobacteria (6 and 10%, respectively), Cytophaga-Flexibacter-Bacteroides (19%), high-G+C-content gram-positive organisms (Actinobacteria; 25%), and low-G+C-content gram-positive organisms (Bacillus and Clostridium; 19%). Twelve percent were identified as chloroplasts. The remaining 9% represented β- and δ-Proteobacteria, Verrucomicrobiales, and candidate divisions. Mixolimnion and oxycline samples had low microbial diversity, with only 9 and 12 distinct phylotypes, respectively, whereas chemocline and monimolimnion samples were more diverse, containing 27 and 25 phylotypes, respectively. The compositions of microbial assemblages from the mixolimnion and oxycline were not significantly different from each other (P = 0.314 and 0.877), but they were significantly different from those of chemocline and monimolimnion assemblages (P < 0.001), and the compositions of chemocline and monimolimnion assemblages were not significantly different from each other (P = 0.006 and 0.124). The populations of sequences retrieved from the mixolimnion and oxycline samples were dominated by sequences related to high-G+C-content gram-positive bacteria (49 and 63%, respectively) distributed in only three distinct phylotypes, while the population of sequences retrieved from the monimolimnion sample was dominated (52%) by sequences related to low-G+C-content gram-positive bacteria distributed in 12 distinct phylotypes. Twelve and 28% of the sequences retrieved from the chemocline sample were also found in the mixolimnion and monimolimnion samples, respectively. None of the sequences retrieved from the monimolimnion sample were found in the mixolimnion or oxycline samples. Elevated diversity in anoxic bottom water samples relative to oxic surface water samples suggests a greater opportunity for niche differentiation in bottom versus surface waters of this lake.     

Artemia monica cyst production and recruitment in Mono Lake,California, USA

Annual egg production was determined for Artemia monica in Mono Lake, California, from 1983 to 1987. Annual oviparous (overwintering cyst) production was 3 and 7 million cysts m^–2 yr^–1 in 1986 and 1987, respectively, as measured by in situ sediment traps. Cyst production for the entire five year period was calculated using Artemia census data and inter-brood duration derived from mixolimnetic temperature. These estimates ranged from 2 to 5 million cysts m^–2 yr^–1. This method underestimated annual production by 30%, when compared to estimates using sediment traps. Cyst production was similar during 1983–1986 and showed a significant increase in 1987, which was due primarily to a larger reproductive population later in the year. Recruitment into the adult populations of the following spring ranged between 1.4 to 3.2%. Overall abundance of this generation reflected the patterns in annual cyst production. Compensatory effects must operate on the second generation of each year, since summer populations were similar in all years despite differences in cyst production.     

Nearshore and pelagic abundances of Artemia monica in Mono Lake,California

The spatial distribution and abundance of the brine shrimp, Artemia monica, in Mono Lake, California were determined during 1982 and 1983. Peak abundances of shrimp occur in midsummer and reach densities of 15–17 individuals l^-1 in the nearshore regions and 6–8 individuals 1^-1 in the pelagic region. The brine shrimp were non-uniformly distributed both vertically and horizontally. The coefficient of variation in shrimp abundance among stations within the nearshore region was similar to that found in the pelagic region. On two of the nine dates, nearshore densities were 3 to 4 times greater than those in the pelagic zone, and on average the brine shrimp appear to be slightly over-dispersed to the nearshore region. However, including nearshore abundances in lakewide estimates will usually result in a change of less than a 10%.     

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

[This corrects the article on p. 614 in vol. 56.].     

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.     

Coupled Arsenotrophy in a Hot Spring Photosynthetic Biofilm at Mono Lake,California

Red-pigmented biofilms grow on rock and cobble surfaces present in anoxic hot springs located on Paoha Island in Mono Lake. The bacterial community was dominated (∼ 85% of 16S rRNA gene clones) by sequences from the photosynthetic Ectothiorhodospira genus. Scraped biofilm materials incubated under anoxic conditions rapidly oxidized As(III) to As(V) in the light via anoxygenic photosynthesis but could also readily reduce As(V) to As(III) in the dark at comparable rates. Back-labeling experiments with ⁷³As(V) demonstrated that reduction to ⁷³As(III) also occurred in the light, thereby illustrating the cooccurrence of these two anaerobic processes as an example of closely coupled arsenotrophy. Oxic biofilms also oxidized As(III) to As(V). Biofilms incubated with [¹⁴C]acetate oxidized the radiolabel to ¹⁴CO₂ in the light but not the dark, indicating a capacity for photoheterotrophy but not chemoheterotrophy. Anoxic, dark-incubated samples demonstrated As(V) reduction linked to additions of hydrogen or sulfide but not acetate. Chemoautotrophy linked to As(V) as measured by dark fixation of [¹⁴C]bicarbonate into cell material was stimulated by either H₂ or HS⁻. Functional genes for the arsenate respiratory reductase (arrA) and arsenic resistance (arsB) were detected in sequenced amplicons of extracted DNA, with about half of the arrA sequences closely related (∼98% translated amino acid identity) to those from the family Ectothiorhodospiraceae. Surprisingly, no authentic PCR products for arsenite oxidase (aoxB) were obtained, despite observing aerobic arsenite oxidation activity. Collectively, these results demonstrate close linkages of these arsenic redox processes occurring within these biofilms.Oxyanions of the group 15 element arsenic, arsenate [As(V)] and arsenite [As(III)], have been known for millennia to be potent poisons. Despite its well-established toxicity to life, the phenomenon of arsenic resistance was discovered whereby some microorganisms maintain an otherwise “normal” existence in the presence of high concentrations of As(V) or As(III) (17, 29, 31). More recently it has become recognized that certain representatives from the bacterial and archaeal domains can actually exploit the electrochemical potential of the As(V)/As(III) redox couple (+130 mV) to gain energy for growth. This can be achieved either by employing As(III) as an autotrophic electron donor or by using As(V) as a respiratory electron acceptor (18, 21, 34). The latter phenomenon, although most commonly associated with chemoheterotrophy, can also employ inorganic substances like sulfide or H₂. Indeed, As(V)-respiring anaerobes displaying a capacity for chemoautotrophy with these electron donors have been isolated and described (5, 7, 16). We recently reported that photoautotrophy is supported by As(III) in anoxic biofilms located in hot springs on Paoha Island in Mono Lake, CA (15). This process represented a novel means of As(III) oxidation achieved via anoxygenic photosynthesis occurring in certain photosynthetic bacteria (i.e., Ectothiorhodospira) and possibly within some cyanobacteria as well (e.g., “Oscillatoria”).Whether or not a microbial habitat is overtly oxic or anoxic, or temporally shifts between these two states over a diel cycle, critical energy linkages between aerobes and anaerobes have long been known for the biogeochemical cycles of key elements, such as sulfur, iron, and nitrogen. Most prominently studied is the case of nitrogen, whereby an ecological coupling exists between the processes of nitrification and denitrification (9, 10, 28). The former process provides energy to aerobic nitrifiers, while the latter process consumes the nitrate produced by this reaction, thereby meeting the energy needs of the denitrifiers.For arsenic, the detection of both As(III) oxidation and As(V) reduction in oxic and anoxic incubations of freshly collected periphyton suggested that an analogous coupled process may also occur for this element (12). Similarly, several uncontaminated soils in Japan displayed a capacity for either As(V) reduction or As(III) oxidation upon arsenic oxyanion amendment and whether they were incubated under oxic or anoxic conditions (39). A defined coculture consisting of an aerobic As(III) oxidizer (strain OL1) and an anaerobic As(V) respirer (strain Y5) was shown to function in this fashion under manipulated laboratory conditions of oxygen tension (26). We pursued the phenomenon of coupled arsenic metabolism further by using materials collected from the hot spring biofilms in Mono Lake, but we focused on examination of the cycling of arsenic under anoxic conditions.In this paper we report results obtained by manipulated incubations of red-pigmented biofilms found in the hot springs of Paoha Island. Preliminary community characterizations of these biofilms show that they are dominated by Bacteria from the genus Ectothiorhodospira but also harbor an assemblage of Archaea related to the Halobacteriacaea. Incubation results have demonstrated the presence of the following arsenic metabolic activities: respiratory As(V) reduction, photosynthetic anaerobic As(III) oxidation, and aerobic As(III) oxidation, along with the ecophysiological conditions under which they occur. Surprisingly, we were unable to obtain authentic PCR products for arsenite oxidase genes (aoxB), despite observing aerobic As(III) oxidation activity. These biofilms serve as a model system for how anaerobic cycling of arsenic can be sustained with oxidation of As(III) by anoxygenic photosynthesis coupled to regeneration of this electron donor via dissimilatory As(V) reduction. The significance that such a light-driven anaerobic ecosystem may have played in the Archean Earth is discussed.     

Distribution of RuBisCO Genotypes along a Redox Gradient in Mono Lake, California

Partial sequences of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) (EC 4.1.1.39) genes were retrieved from samples taken along a redox gradient in alkaline, hypersaline Mono Lake, Calif. The form I gene (cbbL) was found in all samples, whereas form II (cbbM) was not retrieved from any of the samples. None of the RuBisCO sequences we obtained were closely related (nucleotide similarity, <90%) to sequences in the database. Some could be attributed to organisms isolated from the lake (Cyanobium) or appearing in enrichment cultures. Most (52%) of the sequences fell into in one clade, containing sequences that were identical to sequences retrieved from an enrichment culture grown with nitrate and sulfide, and another clade contained sequences identical to those retrieved from an arsenate-reducing, sulfide-oxidizing enrichment.     

In situ hatching of Artemia monica cysts in hypersaline Mono Lake,California

Two emergence trap designs were tested in Mono Lake, California, to measure in situ hatching of Artemia monica cysts on the lake bottom. One design incorporated a removable sample bottle; the other had a catch tube which was pumped from the surface. Both traps rested on the bottom and had a narrow gap between the collecting funnel and bottom flange to allow the chemical conditions within the trap to be similar to those outside. This gap was open during April and May but, because some animals entered from outside the area enclosed by the trap, the gap was covered with 400 µm or 800 µm screen during June and July. The two trap types without screens sampled a station in oxic water 7 m deep similarly in April and May 1985. Mean daily hatching rates from April to May 1985 ranged from 720 to 25 340 shrimp m^-2 day^-1. In contrast, mean daily hatching rates during the same period at a station in anoxic water 21 m deep were from 3 to 138 shrimp m^-2 day^-1. June and July hatching rates in the shallow station were lower than in the spring, usually less than 1000 shrimp m^-2 day^-1.     

Distribution of RuBisCO genotypes along a redox gradient in Mono Lake, California

Partial sequences of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) (EC 4.1.1.39) genes were retrieved from samples taken along a redox gradient in alkaline, hypersaline Mono Lake, Calif. The form I gene (cbbL) was found in all samples, whereas form II (cbbM) was not retrieved from any of the samples. None of the RuBisCO sequences we obtained were closely related (nucleotide similarity, <90%) to sequences in the database. Some could be attributed to organisms isolated from the lake (Cyanobium) or appearing in enrichment cultures. Most (52%) of the sequences fell into in one clade, containing sequences that were identical to sequences retrieved from an enrichment culture grown with nitrate and sulfide, and another clade contained sequences identical to those retrieved from an arsenate-reducing, sulfide-oxidizing enrichment.