Microbial amino acid composition (AA) reflects adaptive strategies of cellular and molecular regulations such as a high proportion of acidic AAs, including glutamic and aspartic acids in alkaliphiles. It remains understudied how microbial AA content is linked to their pH adaptation especially in natural environments. Here we examined prokaryotic communities and their AA composition of genes with metagenomics for 39 water and sediments of East African lakes along a gradient of pH spanning from 7.2 to 10.1. We found that Shannon diversity declined with the increasing pH and that species abundance were either positively or negatively associated with pH, indicating their distinct habitat preference in lakes. Microbial communities showed higher acidic proteomes in alkaline than neutral lakes. Species acidic proteomes were also positively correlated with their pH preference, which was consistent across major bacterial lineages. These results suggest selective pressure associated with high pH likely shape microbial amino acid composition both at the species and community levels. Comparative genome analyses further revealed that alkaliphilic microbes contained more functional genes with higher acidic AAs when compared to those in neutral conditions. These traits included genes encoding diverse classes of cation transmembrane transporters, antiporters, and compatible solute transporters, which are involved in cytoplasmic pH homeostasis and osmotic stress defense under high pH conditions. Our results provide the field evidence for the strong relationship between prokaryotic AA composition and their habitat preference and highlight amino acid optimization as strategies for environmental adaptation.

The genus Natronospira is represented by a single species of extremely salt-tolerant aerobic alkaliphilic proteolytic bacterium, isolated from hypersaline soda lakes. When cells of Gram-positive cocci were used as a substrate instead of proteins at extremely haloalkaline conditions, two new members of this genus were enriched and isolated in pure culture from the same sites. Strains AB-CW1 and AB-CW4 are obligate aerobic heterotrophic proteolytic bacteria able to feed on both live and dead cells of staphylococci and a range of proteins and peptides. Similar to the type species, N. proteinivora, the isolates are extremely salt-tolerant obligate alkaliphiles. However, N. proteinivora was unable to use bacterial cells as a substrate. Electron microscopy showed direct contact between the prey and predator cells. Functional analysis of the AB-CW1 and AB-CW4 genomes identified two sets of genes coding for extracellular enzymes potentially involved in the predation and proteolysis, respectively. The first set includes several copies of lysozyme-like GH23 peptidoglycan-lyase and murein-specific M23 [Zn]-di-peptidase enabling the cell wall degradation. The second set features multiple copies of secreted serine and metallopeptidases apparently allowing for the strong proteolytic phenotype. Phylogenomic analysis placed the isolates into the genus Natronospira as two novel species members, and furthermore indicated that this genus forms a deep-branching lineage of a new family (Natronospiraceae) and order (Natronospirales) within the class Gammaproteobacteria. On the basis of distinct phenotypic and genomic properties, strain AB-CW1T (JCM 335396 = UQM 41579) is proposed to be classified as Natronospira elongata sp. nov., and AB-CW4T (JCM 335397 = UQM 41580) as Natronospira bacteriovora sp. nov.

Beta-glucosidases catalyze the hydrolysis of the glycosidic bonds of cellobiose, producing glucose, which is a rate-limiting step in cellulose biomass degradation. In industrial processes, β-glucosidases that are tolerant to glucose and stable under harsh industrial reaction conditions are required for efficient cellulose hydrolysis. In this study, we report the molecular cloning, Escherichia coli expression, and functional characterization of a β-glucosidase from the gene, CelGH3_f17, identified from metagenomics libraries of an Ethiopian soda lake. The CelGH3_f17 gene sequence contains a glycoside hydrolase family 3 catalytic domain (GH3). The heterologous expressed and purified enzyme exhibited optimal activity at 50 °C and pH 8.5. In addition, supplementation of 1 M salt and 300 mM glucose enhanced the β-glucosidase activity. Most of the metal ions and organic solvents tested did not affect the β-glucosidase activity. However, Cu2+ and Mn2+ ions, Mercaptoethanol and Triton X-100 reduce the activity of the enzyme. The studied β-glucosidase enzyme has multiple industrially desirable properties including thermostability, and alkaline, salt, and glucose tolerance.

Soda lakes are some of the most productive aquatic ecosystems.1 Their alkaline-saline waters sustain unique phytoplankton communities2,3 and provide vital habitats for highly specialized biodiversity including invertebrates, endemic fish species, and Lesser Flamingos (Phoeniconaias minor).1,4 More than three-quarters of Lesser Flamingos inhabit the soda lakes of East Africa5; however, populations are in decline.6 Declines could be attributed to their highly specialized diet of cyanobacteria7 and dependence on a network of soda lake feeding habitats that are highly sensitive to climate fluctuations and catchment degradation.8,9,10,11,12 However, changing habitat availability has not been assessed due to a lack of in situ water quality and hydrology data and the irregular monitoring of these waterbodies.13 Here, we combine satellite Earth observations and Lesser Flamingo abundance observations to quantify spatial and temporal trends in productivity and ecosystem health over multiple decades at 22 soda lakes across East Africa. We found that Lesser Flamingo distributions are best explained by phytoplankton biomass, an indicator of food availability. However, timeseries analyses revealed significant declines in phytoplankton biomass from 1999 to 2022, most likely driven by substantial rises in lake water levels. Declining productivity has reduced the availability of healthy soda lake ecosystems, most notably in equatorial Kenya and northern Tanzania. Our results highlight the increasing vulnerability of Lesser Flamingos and other soda lake biodiversity in East Africa, particularly with increased rainfall predicted under climate change.14,15,16 Without improved lake monitoring and catchment management practices, soda lake ecosystems could be pushed beyond their environmental tolerances. VIDEO ABSTRACT.

A variety of lakes located in the dry steppe area of southwestern Siberia are exposed to rapid climatic changes, including intra-century cycles with alternating dry and wet phases driven by solar activity. As a result, the salt lakes of that region experience significant fluctuations in water level and salinity, which have an essential impact on the indigenous microbial communities. But there are few microbiological studies that have analyzed this impact, despite its importance for understanding the functioning of regional water ecosystems. This work is a retrospective study aimed at analyzing how solar activity-related changes in hydrological regime affect phototrophic microbial communities using the example of the shallow soda lake Tanatar VI, located in the Kulunda steppe (Altai Region, Russia, southwestern Siberia). The main approach used in this study was the comparison of hydrochemical and microscopic data obtained during annual field work with satellite and solar activity data for the 12-year observation period (2011-2022). The occurrence of 33 morphotypes of cyanobacteria, two key morphotypes of chlorophytes, and four morphotypes of anoxygenic phototrophic bacteria was analyzed due to their easily recognizable morphology. During the study period, the lake surface changed threefold and the salinity changed by more than an order of magnitude, which strongly correlated with the phases of the solar activity cycles. The periods of high (2011-2014; 100-250 g/L), medium (2015-2016; 60 g/L), extremely low (2017-2020; 13-16 g/L), and low (2021-2022; 23-34 g/L) salinity with unique biodiversity of phototrophic communities were distinguished. This study shows that solar activity cycles determine the dynamics of the total salinity of a southwestern Siberian soda lake, which in turn determines the communities and microorganisms that will occur in the lake, ultimately leading to cyclical changes in alternative states of the ecosystem (dynamic stability).

Haloalkaliphilic chemolithoautotrophic sulfur-oxidizing bacteria belonging to the genus Thioalkalivibrio are highly abundant in microbial communities found in soda lakes and dominant in full-scale bioreactors removing sulfide from industrial waste gases. Despite certain soda lakes being remote and unaffected by anthropogenic activities, haloalkaliphilic microorganisms, including Thioalkalivibrio strains, possess various antibiotic resistance genes. In this study, we investigated the impact of the antibiotic ampicillin on a co-culture of two Thioalkalivibrio species, Tv. thiocyanoxidans ARh2T and Tv. versutus AL2T, both experimentally and through in silico analysis of antibiotic resistance. Cell growth dynamics were monitored over time at increasing ampicillin concentrations using rep- and qPCR. Within ten days after the addition of ampicillin, the co-culture transitioned from a Tv. thiocyanoxidans ARh2T-dominated to a stable Tv. versutus AL2T-dominated culture. This shift was attributed to Tv. versutus AL2T displaying a lower susceptibility to ampicillin, making it more competitive. These results emphasize the potential implications of antibiotic pressure on microbial communities, where a resistant species can outcompete a stable co-culture. This study presents the first evidence of such dynamics in haloalkaliphilic chemolithoautotrophs. By understanding the antibiotic resistance and the competitive dynamics of haloalkaliphilic bacteria like Thioalkalivibrio, we can gain insights into their behaviour and stress response.

Haloalkaliphilic microorganisms are double extremophiles functioning optimally at high salinity and pH. Their typical habitats are soda lakes, geologically ancient yet widespread ecosystems supposed to harbor relict microbial communities. We compared metabolic features and their determinants in two strains of the natronophilic species Dethiobacter alkaliphilus, the only cultured representative of the class \"Dethiobacteria\" (Bacillota). The strains of D. alkaliphilus were previously isolated from geographically remote Mongolian and Kenyan soda lakes. The type strain AHT1T was described as a facultative chemolithoautotrophic sulfidogen reducing or disproportionating sulfur or thiosulfate, while strain Z-1002 was isolated as a chemolithoautotrophic iron reducer. Here, we uncovered the iron reducing ability of strain AHT1T and the ability of strain Z-1002 for thiosulfate reduction and anaerobic Fe(II) oxidation. Key catabolic processes sustaining the growth of both D. alkaliphilus strains appeared to fit the geochemical settings of two contrasting natural alkaline environments, sulfur-enriched soda lakes and iron-enriched serpentinites. This hypothesis was supported by a meta-analysis of Dethiobacterial genomes and by the enrichment of a novel phylotype from a subsurface alkaline aquifer under Fe(III)-reducing conditions. Genome analysis revealed multiheme c-type cytochromes to be the most probable determinants of iron and sulfur redox transformations in D. alkaliphilus. Phylogeny reconstruction showed that all the respiratory processes in this organism are likely provided by evolutionarily related early forms of unconventional octaheme tetrathionate and sulfite reductases and their structural analogs, OmhA/OcwA Fe(III)-reductases. Several phylogenetically related determinants of anaerobic Fe(II) oxidation were identified in the Z-1002 genome, and the oxidation process was experimentally demonstrated. Proteomic profiling revealed two distinct sets of multiheme cytochromes upregulated in iron(III)- or thiosulfate-respiring cells and the cytochromes peculiar for Fe(II) oxidizing cells. We suggest that maintaining high variation in multiheme cytochromes is an effective adaptive strategy to occupy geochemically contrasting alkaline environments. We propose that sulfur-enriched soda lakes could be secondary habitats for D. alkaliphilus compared to Fe-rich serpentinites, and that the ongoing evolution of Dethiobacterales could retrace the evolutionary path that may have occurred in prokaryotes at a turning point in the biosphere\'s history, when the intensification of the sulfur cycle outweighed the global significance of the iron cycle.

Soda lakes are productive natural ecosystems with substantial economic and non-economic values. Currently, they are facing considerable environmental threats that can lead to further degradation. The objective of this study was to investigate comparative spatiotemporal variations of physicochemical properties of four Ethiopian soda lakes in comparison with their historical data. Central (open-water) sampling sites were selected from four Ethiopian soda lakes: Arenguade, Beseka, Chittu and Shala. Water samples were collected from open sampling stations from January to December 2020 and analyzed at Limnology laboratory of Addis Ababa University. The geographical position of each lake was determined by means of Global Positioning System (GPS). All physicochemical factors exhibited significant differences between seasons (ANOVA, P < 0.05), except salinity in Lake Shala. The concentrations of physicochemical parameters were generally high during the dry seasons in the studied lakes due to the low incidence of rainfall, caused by recurrent drought, resulting in higher evapotranspiration rates as they are characterized by a long dry season. Lakes Arenguade and Beseka showed considerable decrease in conductivity, alkalinity and salinity, compared to data from the 1960s and 1990s, which might be attributed to dilution effect. The same parameters show slightly increasing trend in Lake Arenguade which might be due to high evaporation rate. In general, the physicochemical parameters of the study lakes showed temporal variations, which could be attributed to the dilution effect, evaporation, and hydrological characteristics of the Ethiopian Rift Valley. In the face of climate change and recurring droughts, in the Ethiopian Rift Valley, the outcomes of this study might be used as input for the long-term planning, of water resources management and devising mitigation strategies.

Natural, inland alkaline soda waters form a particular type of saline waters, characterized by a permanent alkaline chemical property. In many cases only the total alkalinity by methyl-orange titration is reported, without phenolphthalein titration. Therefore, a reliable estimation of carbonates from total alkalinity is essential for a precise scientific chemical classification. The concentration of bicarbonate [HCO3 ‒] can be reliably estimated in waters using the Advanced Speciation Method (ASM) if methyl-orange total alkalinity titration and pH data are available, while the concentration of carbonate [CO3 2‒] is not reliably estimated by the ASM when interfering factors with acid/base properties (e.g., phosphate, silicate, ammonia, etc.) are present in significant concentrations in natural waters. Therefore, here I present and prove an experimental polynomial function for carbonate estimation with the following equation based on the concentration of bicarbonate: [CO3 2‒] = -2.878E-7 ± 5.438E-8 × [HCO3 ‒]2 + 0.069±0.003 × [HCO3 ‒] This Boros\'s carbonate estimation method can contribute to a more efficient evaluation of field water samples with several analytical difficulties.•Bicarbonate can be reliably estimated using the Advanced Speciation Method (ASM).•Estimation of the carbonate concentration using ASM in the presence of interfering acid/base factors in alkaline waters.•Experimental polynomial function for reliable carbonate estimation in alkaline soda waters.

The occurrence of high arsenic concentrations (up to 3000 μg L-1) in water of soda lakes of the Pantanal wetland is a remarkable case of natural arsenic contamination in South America. However, little is known about arsenic speciation in this environment, particularly regarding speciation changes related to lake trophic status and seasonal variations. To fill this gap, arsenic speciation analysis was carried out in surface (SW) and subsurface (SSW) waters sampled in five soda lakes with different eutrophication status, in two dry and one wet season. As(V) was the dominant species in these waters, while As(III), DMA, MMA and likely complex organic species were present in lower amounts. The results allow to conclude that the arsenic speciation in SW and SSW varies seasonally according to the regional wet or dry periods and lake water levels. In eutrophic turbid and in oligotrophic vegetated soda lakes, arsenic speciation was also characterized by spatial differences between edge and center or between the SW and SSW. Cyanobacteria or macrophytes/algae are involved in arsenic biotransformation in soda lakes through its metabolic and detoxification processes. Significant variation in surface water arsenic speciation occurs as a result of seasonal primary production fluctuation or water arsenic concentration changes in the soda lakes, increasing organoarsenics in dry periods, whereas in flood periods, As(V) prevails. Spatial distribution of arsenic species is significantly impacted by biogeochemical conditions at the water/sediment interface in soda lakes.