Hot Spring Samples Research Articles

Migration and controlling factors of rare earth elements in geothermal systems on the southern Tibetan Plateau

Rare earth elements (REE) serve as effective tracers of water-rock interactions, a crucial process in the evolution of geothermal systems. However, the geochemical behavior of REE during their migration within geothermal systems, particularly the fractionation mechanism, remains poorly understood. In this study, we focus on the geochemical characteristics of REE in hot springs, host rocks, and sinters from various hot spring systems in the Yadong-Gulu rift (YGR), the largest extensional rift in the southern Tibetan Plateau. The objective is to explore the factors that control REE concentrations in hot springs and elucidate the fractionation of REE during their migration. The concentrations of REE are significantly influenced by pH, Fe oxides (oxyhydroxides), and HCO3− concentrations in central and southern hot springs, whereas high reservoir temperatures contribute to higher ΣREE concentrations in northern hyperthermal springs. REE speciation calculations show that LnO2− and LnO2H are dominant complexes in northern alkaline hot springs, while carbonate complexes (LnCO3+ and Ln(CO3)2−) prevail in central and southern spring samples. Additionally, weakly acidic hot springs exhibit a prevalence of Ln3+, LnF2+, and LnSO4+ species. The chondrite-normalized REE patterns of all hot spring samples display enrichment in MREE, which is significantly distinct from the patterns observed in host rocks and sinters enriched in LREE. The fractionation of REE in the YGR hot spring systems is primarily controlled by the following processes: (1) Temperature, pH, and relative abundance of complexing agents influence the complexation of REE and further regulate their fractionation in hot springs; (2) During water-rock interaction, preferential leaching of HREE and MREE from host rocks can lead to their enrichment in alkaline hot springs. The dissolution of iron-rich sediments plays a significant role in generating the distinctive MREE bulge pattern in acid-neutral hot springs compared to host rocks; (3) The higher stability of MREE and HREE complexes in water, as well as the preferential coprecipitation of unstable LnCO3+ complexes formed by LREE with carbonates, are key factors responsible for LREE enrichment in sinters.

Multi-omics insights into the function and evolution of sodium benzoate biodegradation pathway in Benzoatithermus flavus gen. nov., sp. nov. from hot spring

Biodegradation stands as an eco-friendly and effective approach for organic contaminant remediation. However, research on microorganisms degrading sodium benzoate contaminants in extreme environments remains limited. In this study, we report to display the isolation of a novel hot spring enriched cultures with sodium benzoate (400 mg/L) as the sole carbon source. The results revealed that the phylum Pseudomonadota was the potential sodium benzoate degrader and a novel genus within the family Geminicoccaceae of this phylum. The isolated strain was named Benzoatithermus flavus SYSU G07066T and was isolated from HNT-2 hot spring samples. Genomic analysis revealed that SYSU G07066T carried benABC genes and physiological experiments indicated the ability to utilize sodium benzoate as a sole carbon source for growth, which was further confirmed by transcriptomic data with expression of benABC. Phylogenetic analysis suggested that Horizontal Gene Transfer (HGT) plays a significant role in acquiring sodium benzoate degradation capability among prokaryotes, and SYSU G07066T might have acquired benABC genes through HGT from the family Acetobacteraceae. The discovery of the first microorganism with sodium benzoate degradation function from a hot spring enhances our understanding of the diverse functions within the family Geminicoccaceae. This study unearths the first novel genus capable of efficiently degrading sodium benzoate and its evolution history at high temperatures, holding promising industrial applications, and provides a new perspective for further exploring the application potential of hot spring “microbial dark matter”.

Hydrogeochemistry and Isotope Characteristics of the Hot Springs in the Wapsalit Area, Buru Regency, Maluku Province, Indonesia

Wapsalit is one of the geothermal areas that is located in Buru Regency, Maluku Province. Wapsalit area has several geothermal manifestations like hot springs, altered rocks, silica sinter and many more. This research aims to determine the characteristics and origin of the fluid in the research area. This research takes 5 samples of hot springs named FATH-1, FATH-2, FATH-3, FATH-4 and FATH-5. Based on the results of chemical analysis, it shows that the 5 samples belong to the bicarbonate fluid type. FATH-1, FATH-3 and FATH-5 belong to the partial equilibrium zone, FATH-2 and FATH-4 belong to the immature water zone. Based on isotope 18O and D analysis, the 5 hot spring samples originated from meteoric water and the fluids interacting with the rocks before heading to the surface.

Genomic insight and physiological characterization of thermoacidophilic Alicyclobacillus isolated from Yellowstone National Park.

Alicyclobacillus has been isolated from extreme environments such as hot springs, volcanoes, as well as pasteurized acidic beverages, because it can tolerate extreme temperatures and acidity. In our previous study, Alicyclobacillus was isolated during the enrichment of methane oxidizing bacteria from Yellowstone Hot Spring samples. Physiological characterization and genomic exploration of two new Alicyclobacillus isolates, AL01A and AL05G, are the main focus of this study to identify their potential relationships with a thermoacidophilic methanotroph (Methylacidiphilum) isolated from the same hot spring sediments. In the present study, both Alicyclobacillus isolates showed optimal growth at pH 3.5 and 55°C, and contain ω-alicyclic fatty acids as a major lipid (ca. 60%) in the bacterial membrane. Genomic analysis of these strains revealed specific genes and pathways that the methanotroph genome does not have in the intermediary carbon metabolism pathway such as serC (phosphoserine aminotransferase), comA (phosphosulfolactate synthase), and DAK (glycerone kinase). Both Alicyclobacillus strains were also found to contain transporter systems for extracellular sulfate (ABC transporter), suggesting that they could play an important role in sulfur metabolism in this extreme environment. Genomic analysis of vitamin metabolism revealed Alicyclobacillus and Methylacidiphilum are able to complement each other's nutritional deficiencies, resulting in a mutually beneficial relationship, especially in vitamin B1(thiamin), B3 (niacin), and B7 (biotin) metabolism. These findings provide insights into the role of Alicyclobacillus isolates in geothermal environments and their unique metabolic adaptations to these environments.

Use the hot spring’s Chloro-Alkaline Index (CAI) for the low enthalpy prospect of the Rawadanau geothermal field

The emergence of hot springs along the volcanic shelf in Indonesia is a feature of geothermal manifestations. The geothermal system is affected by heat sources, reservoir rocks, cap rocks, and meteoric water that fills and escapes as hot springs through fractures. The Rawadanau geothermal field in Serang Banten is low enthalpy prospects. Geothermal maintenance is defined by the chemical composition of the hot springs, one of which can be determined by the ion exchange rate, namely the chloro-alkaline index (CAI) method. There are 16 hot spring samples, with temperatures (31.2 - 54.6 °C), pH (6.2 – 7.3), and EC (1190 – 4460 μS/cm), respectively. The water types encountered are Na-Cl in the southern and Mg-Ca-HCO3, and Na-Mg-Ca-HCO3-Cl in the northern and western, respectively. Generally, samples taken in the rainy show negative CAI and positive CAI in the dry, while Cl-type is positive in two seasons. Based on water type and CAI show that the Na-Cl type tends to be positive in the two seasons. While the Ca-Mg-HCO3 type is positive in the dry and negative in the rainy, and the Mixed-type shows a negative CAI. The results of this study provide information for the maintenance of geothermal fields as they are developed.

Analysis of Trace Mineral Elements and Microorganisms in Two Hot Springs in Jazan Region of Saudi Arabia

Thermal springs have hydrothermal or volcanic activity, which causes the water to boil up and dissolve any metal that may be present there. It helps in recovery to reduce stress, promote sleep, boost blood circulation, relieve pain and heal skin problems. However, they consist of some heavy and trace metals and some non-metals, such as Mg, Mo, Na, As, Hg, Cd, Ni, Fe, Cu, Mn, Zn, Co, Ti, Cr and Pb are present. These hot springs are also known to exhibit natural microbiota, which is helpful in maintaining the public health and preventing consumer outbreaks with the advancement of research, microbiological monitoring of hot spring water can be done by utilizing high throughput sequencing. Taking these facts in concern, the present work, hot spring samples were collected from different locations of Al-Harth and Alarda hot springs, Jazan region, Saudi Arabia. The samples collected were analyzed for trace minerals and microbiological activity. These samples were found to exhibit the temperature range from 60 to 70 ºC and an average pH of 6.5 to 8.5. Almost all the samples had shown the presence of trace elements with electrical conductivity and total dissolved solids (TDS) ranged from 0.02 to 0.045 dms and 27 to 43 mg/L, respectively. Scattergun metagenomic analysis was performed to determine the microbiological content. The preliminary study reports the microbiome (bacteria, archea and viruses) of alarath hot spring water employing whole-genome high-throughput sequencing and bioinformatics. Bacterial species detected were members of the phyla 64% Proteobacteria, 20% Actinobacteria, 6.3% unclassified, 2.2% Deinococcus thermus, 1.4 Firmcutes and 1.6% others. Fecal indicator bacteria, i.e. Escherichia coli or enterococci, were not detected in any samples examined in this study. Both archaeal DNA (0.4%) and viral DNA (0.009%) were found.

Determination of arsenic and antimony by atmospheric pressure glow discharge portable emission spectrometer with solid acid hydride generation

In this work, the use of solid acid hydride generation (HG) for the simultaneous determination of As and Sb in environmental water samples by miniaturized atmospheric pressure glow discharge plasma optical emission spectrometer (APGD-OES) was evaluated. It was found that HG with the use of sulfamic acid tablets can achieve comparable sensitivity with conventional HCl-KBH4 for As and Sb determination by APGD-OES, which avoids the safety issue encountered in the use of hydrochloric acid and greatly facilitates field analysis. Under optimal conditions, the obtained detection limits (LODs) are as low as 1.26 μg L−1 for As and 0.72 μg L−1 for Sb, respectively. Moreover, it offers good repeatability, where the relative standard deviations (RSDs) are less than 2.5%. The accuracy of the proposed method was validated by the analysis of simulated natural water reference samples. Furthermore, it has been successfully applied to the determination of tap water, pond water, and hot spring samples. All these results clearly demonstrate the proposed simple, cost-effective and field-deployable method provides a promising approach for the field analysis of As and Sb.

Methanogens-Driven Arsenic Methylation Preceding Formation of Methylated Thioarsenates in Sulfide-Rich Hot Springs

Hot springs represent a major source of arsenic release into the environment. Speciation is typically reported to be dominated by arsenite, arsenate, and inorganic thiolated arsenates. Much less is known about the relevance and formation of methylated thioarsenates, a group with species of high mobility and toxicity. In hot spring samples taken from the Tengchong volcanic region in China, methylated thioarsenates contributed up to 13% to total arsenic. Enrichment cultures were obtained from the corresponding sediment samples and incubated to assess their capability to convert arsenite into methylated thioarsenates over time and in the presence of different microbial inhibitors. In contrast to observations in other environmental systems (e.g., paddy soils), there was no solid evidence, supporting that the sulfate-reducing bacteria contributed to the arsenic methylation. Methanosarcina, the sole genus of methanogens detected in the enrichment cultures, as well as Methanosarcina thermophila TM-1, a pure strain within the genus, did methylate arsenic. We propose that methylated thioarsenates in a typical sulfide-rich hot spring environment like Tengchong form via a combination of biotic arsenic methylation driven by thermophilic methanogens and arsenic thiolation with either geogenic sulfide or sulfide produced by sulfate-reducing bacteria.

Microbial ecology of a shallow alkaline hydrothermal vent: Strýtan Hydrothermal Field, Eyjafördur, northern Iceland.

Strýtan Hydrothermal Field (SHF) is a submarine system located in Eyjafördur in northern Iceland composed of two main vents: Big Strýtan and Arnarnesstrýtan. The vents are shallow, ranging from 16 to 70 m water depth, and vent high pH (up to 10.2), moderate temperature (T max ∼70°C), anoxic, fresh fluids elevated in dissolved silica, with slightly elevated concentrations of hydrogen and methane. In contrast to other alkaline hydrothermal vents, SHF is unique because it is hosted in basalt and therefore the high pH is not created by serpentinization. While previous studies have assessed the geology and geochemistry of this site, the microbial diversity of SHF has not been explored in detail. Here we present a microbial diversity survey of the actively venting fluids and chimneys from Big Strýtan and Arnarnesstrýtan, using 16S rRNA gene amplicon sequencing. Community members from the vent fluids are mostly aerobic heterotrophic bacteria; however, within the chimneys oxic, low oxygen, and anoxic habitats could be distinguished, where taxa putatively capable of acetogenesis, sulfur-cycling, and hydrogen metabolism were observed. Very few archaea were observed in the samples. The inhabitants of SHF are more similar to terrestrial hot spring samples than other marine sites. It has been hypothesized that life on Earth (and elsewhere in the solar system) could have originated in an alkaline hydrothermal system, however all other studied alkaline submarine hydrothermal systems to date are fueled by serpentinization. SHF adds to our understandings of hydrothermal vents in relationship to microbial diversity, evolution, and possibly the origin of life.

STUDI KIMIA FISIK FLUIDA PADA MANIFESTASI PANAS BUMI DANAU LINOW

Lake Linow is one of the volcanic lakes that has geothermal activity characterized by geothermal manifestations in the form of hot springs that appear on the surface. The purpose of this research is to study the type of fluid and estimate the reservoir temperature. The methods used in this research are ion analysis of fluid content using spectrometry, pH, and physical analysis of fluids in the form of physical appearance manifestations of hot springs. The results showed that the hot springs in Lake Linow are immature water types with a high percentage of Mg concentration. It is interpreted that there is a process of dilution with other elements and mixing with meteoric water which is rich in Mg elements. Calculations using water geothermometer equations on hot spring samples in the area are estimated to have a reservoir temperature of 92.85°C-127.9°C

Geothermal Fluid Characterisics based on Geochemical Analysis of Hot Water in The Wawolesea Area, North Konawe Regency Southeast Sulawesi Province

North Konawe Regency is an area with interesting geothermal potential to be studied. The observed geothermal potential manifestation is located in the Wawolesea area through the collection and analysis of three hot spring samples. This study aims to determine the characteristics of geothermal fluids, namely the type and origin of the fluid by conducting geochemical analysis on samples of hot springs in the study area which further shows the content of elements and chemical compounds and their respective concentrations. The analysis in this research is Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) to obtain B and Li concentrations, conductivity meter to obtain Cl concentration, visible spectroscopy method for SO4 concentration and HCO3 concentration measured by acid-base titration. The results showed that the type of hot springs in the study area was included in the Chloride type with a percentage of 81.8156%-83.3976% with a pH value of 6.7-7.4 and a hot spring temperature of 50-54°C then plotted into trilinear diagram. The Cl-Li-B Ternanry diagram shows that the source of the fluid is in a reservoir that has the same rock structure and comes from the old hydrothermal system. Based on the results of the analysis of fluid characteristics obtained from the research, it can be concluded that the hot springs of the Wawolesea Region are an indicator of the presence of geothermal sources and further research is needed on the magnitude of the geothermal potential.

Influence of Metal Ions on Model Protoamphiphilic Vesicular Systems: Insights from Laboratory and Analogue Studies.

Metal ions strongly affect the self-assembly and stability of membranes composed of prebiotically relevant amphiphiles (protoamphiphiles). Therefore, evaluating the behavior of such amphiphiles in the presence of ions is a crucial step towards assessing their potential as model protocell compartments. We have recently reported vesicle formation by N-acyl amino acids (NAAs), an interesting class of protoamphiphiles containing an amino acid linked to a fatty acid via an amide linkage. Herein, we explore the effect of ions on the self-assembly and stability of model N-oleoyl glycine (NOG)-based membranes. Microscopic analysis showed that the blended membranes of NOG and Glycerol 1-monooleate (GMO) were more stable than pure NOG vesicles, both in the presence of monovalent and divalent cations, with the overall vesicle stability being 100-fold higher in the presence of a monovalent cation. Furthermore, both pure NOG and NOG + GMO mixed systems were able to self-assemble into vesicles in natural water samples containing multiple ions that were collected from active hot spring sites. Our study reveals that several aspects of the metal ion stability of NAA-based membranes are comparable to those of fatty acid-based systems, while also confirming the robustness of compositionally heterogeneous membranes towards high metal ion concentrations. Pertinently, the vesicle formation by NAA-based systems in terrestrial hot spring samples indicates the conduciveness of these low ionic strength freshwater systems for facilitating prebiotic membrane-assembly processes. This further highlights their potential to serve as a plausible niche for the emergence of cellular life on the early Earth.

Fluid Geochemistry within the North China Craton: Spatial Variation and Genesis

The North China Craton (NCC) is a typical representative of the ancient destruction craton. Numerous studies have shown that extensive destruction of the NCC occurred in the east, whereas the western part was only partially modified. The Bohai Bay Basin is in the center of the destruction area in the eastern NCC. Chemical analyses were conducted on 122 hot spring samples taken from the eastern NCC and the Ordos Basin. The δ 2 H and δ 18 O in water, δ 13 C in CO2, and 3He/4He and 4He/20Ne ratios in gases were analyzed in combination with chemical analyses of water in the central and eastern NCC. The results showed an obvious spatial variation in chemical and isotopic compositions of the geofluids in the NCC. The average temperature of spring water in the Trans-North China Block (TNCB) and the Bohai Bay Basin was 80.74°C, far exceeding that of the Ordos Basin of 38.43°C. The average δ D in the Eastern Block (EB) and the TNCB were −79.22‰ and −84.13‰, respectively. The He isotope values in the eastern region (TNCB and EB) ranged from 0.01 to 2.52, and the rate of contribution of the mantle to He ranged from 0 to 31.38%. δ 13 C ranged from −20.7 to −6.4‰ which indicated an organic origin. The chemical compositions of the gases in the EB showed that N2 originated mainly from the atmosphere. The EB showed characteristics of a typical gas subduction zone, whereas the TNCB was found to have relatively small mantle sources. The reservoir temperatures in the Ordos Basin and the eastern NCC (EB and TNCB) calculated by the K-Mg temperature scale were 38.43°C and 80.74°C, respectively. This study demonstrated clear spatial variation in the chemical and isotopic compositions of the geofluids in the NCC, suggesting the presence of geofluids from the magmatic reservoir in the middle-lower crust and that active faults played an important role in the transport of mantle-derived components from the mantle upwards.

Diversity and Distribution of Culturable Thermus Species in Terrestrial Hot Springs of Southwestern Yunnan Province in China

The Yunnan geothermal area has many neutral and alkalescent thermal springs. Members of the genus Thermus have been found in thermal environments. In this study, we attempted to cultivate numerically abundant Thermus species using a variety of different strategies. A total of 223 strains of Thermus-like bacteria were isolated from seventeen hot spring samples of four geothermal regions (Baoshan, Dali, Lincang and Dehong). These strains were classified into two genera, Thermus and Meiothermus, based on 16S rDNA. The optimal isolation temperature of the Thermus genus was 63–70 °C. The highest Thermus diversity was found at 63 °C. Thermus brockianus was a universal culturable bacterium in the four geothermal regions. The principal component analysis (PCA) showed that T. oshimai preferred to inhabit the hot springs of the Baoshan (Bs) geothermal region, and T. amyloliquefaciens dominated the Dali (Dl) geothermal region, whereas T. tengchongensis accumulated in the Lincang (Lc) and Dehong (Dh) geothermal regions. The results suggested that Thermus species had habitat-preferable characteristics among the four geothermal regions. The findings may help identify the niche from which Thermus strains can likely be isolated.

Sasso Pisano Geothermal Field Environment Harbours Diverse Ktedonobacteria Representatives and Illustrates Habitat-Specific Adaptations.

The hydrothermal steam environment of Sasso Pisano (Italy) was selected to investigate the associated microbial community and its metabolic potential. In this context, 16S and 18S rRNA gene partial sequences of thermophilic prokaryotes and eukaryotes inhabiting hot springs and fumaroles as well as mesophilic microbes colonising soil and water were analysed by high-throughput amplicon sequencing. The eukaryotic and prokaryotic communities from hot environments clearly differ from reference microbial communities of colder soil sites, though Ktedonobacteria showed high abundances in various hot spring samples and a few soil samples. This indicates that the hydrothermal steam environments of Sasso Pisano represent not only a vast reservoir of thermophilic but also mesophilic members of this Chloroflexi class. Metabolic functional profiling revealed that the hot spring microbiome exhibits a higher capability to utilise methane and aromatic compounds and is more diverse in its sulphur and nitrogen metabolism than the mesophilic soil microbial consortium. In addition, heavy metal resistance-conferring genes were significantly more abundant in the hot spring microbiome. The eukaryotic diversity at a fumarole indicated high abundances of primary producers (unicellular red algae: Cyanidiales), consumers (Arthropoda: Collembola sp.), and endoparasite Apicomplexa (Gregarina sp.), which helps to hypothesise a simplified food web at this hot and extremely nutrient-deprived acidic environment.

Comparative Metagenomic Analysis of Two Alkaline Hot Springs of Madhya Pradesh, India and Deciphering the Extremophiles for Industrial Enzymes.

Hot springs are considered to be a unique environment with extremophiles, that are sources of industrially important enzymes, and other biotechnological products. The objective of this study was to undertake, analyze, and characterize the microbiome of two major hot springs located in the state of Madhya Pradesh explicitly, Chhoti Anhoni (Hotspring 1), and Badi Anhoni (Hotspring 2) to find out the inhabitant microbial population, and their functional characteristics. The taxonomic analysis of the microbiome of the hot springs revealed the phylum Proteobacteria was the most abundant taxa in both the hot-springs, however, its abundance in hot-spring 1 (~88%) was more than the hot-spring 2 (~52%). The phylum Bacteroides (~10–22%) was found to be the second most abundant group in the hot-springs followed by Spirocheates (~2–11%), Firmicutes (~6–8%), Chloroflexi (1–5%), etc. The functional analysis of the microbiome revealed different features related to several functions including metabolism of organics and degradation of xenobiotic compounds. The functional analysis showed that most of the attributes of the microbiome was related to metabolism, followed by cellular processes and environmental information processing functions. The functional annotation of the microbiomes at KEGG level 3 annotated the sequences into 279 active features that showed variation in abundance between the hot spring samples, where hot-spring 1 was functionally more diverse. Interestingly, the abundance of functional genes from methanogenic bacteria, was higher in the hot-spring 2, which may be related to the relatively higher pH and temperature than Hotspring 1. The study showed the presence of different unassigned bacterial taxa with high abundance which indicates the potential of novel genera or phylotypes. Culturable isolates (28) were bio-prospected for industrially important enzymes including amylase, protease, lipase, gelatinase, pectinase, cellulase, lecithinase, and xylanase. Seven isolates (25%) had shown positive results for all the enzyme activities whereas 23 isolates (82%) produced Protease, 27 isolates (96%) produced lipase, 27 isolates produced amylase, 26 isolates (92%) produced cellulase, 19 isolates (67%) produced pectinase, 19 isolates (67%) could produce lecithinase, and 13 isolates (46%) produced gelatinase. The seven isolates, positive for all the enzymes were analyzed further for quantitative analysis and identified through molecular characterization.

Physicochemical and Microbial Diversity Analyses of Indian Hot Springs.

In the present study, physicochemical and microbial diversity analyses of seven Indian hot springs were performed. The temperature at the sample sites ranged from 32 to 67°C, and pH remained neutral to slightly alkaline. pH and temperature influenced microbial diversity. Culture-independent microbial diversity analysis suggested bacteria as the dominant group (99.3%) when compared with the archaeal group (0.7%). Alpha diversity analysis showed that microbial richness decreased with the increase of temperature, and beta diversity analysis showed clustering based on location. A total of 131 strains (divided into 12 genera and four phyla) were isolated from the hot spring samples. Incubation temperatures of 37 and 45°C and T5 medium were more suitable for bacterial isolation. Some of the isolated strains shared low 16S rRNA gene sequence similarity, suggesting that they may be novel bacterial candidates. Some strains produced thermostable enzymes. Dominant microbial communities were found to be different depending on the culture-dependent and culture-independent methods. Such differences could be attributed to the fact that most microbes in the studied samples were not cultivable under laboratory conditions. Culture-dependent and culture-independent microbial diversities suggest that these springs not only harbor novel microbial candidates but also produce thermostable enzymes, and hence, appropriate methods should be developed to isolate the uncultivated microbial taxa.

Genomic Insights of "Candidatus Nitrosocaldaceae" Based on Nine New Metagenome-Assembled Genomes, Including "Candidatus Nitrosothermus" Gen Nov. and Two New Species of "Candidatus Nitrosocaldus".

“Candidatus Nitrosocaldaceae” are globally distributed in neutral or slightly alkaline hot springs and geothermally heated soils. Despite their essential role in the nitrogen cycle in high-temperature ecosystems, they remain poorly understood because they have never been isolated in pure culture, and very few genomes are available. In the present study, a metagenomics approach was employed to obtain “Ca. Nitrosocaldaceae” metagenomic-assembled genomes (MAGs) from hot spring samples collected from India and China. Phylogenomic analysis placed these MAGs within “Ca. Nitrosocaldaceae.” Average nucleotide identity and average amino acid identity analysis suggested the new MAGs represent two novel species of “Candidatus Nitrosocaldus” and a novel genus, herein proposed as “Candidatus Nitrosothermus.” Key genes responsible for chemolithotrophic ammonia oxidation and a thaumarchaeal 3HP/4HB cycle were detected in all MAGs. Furthermore, genes coding for urea degradation were only present in “Ca. Nitrosocaldus,” while biosynthesis of the vitamins, biotin, cobalamin, and riboflavin were detected in almost all MAGs. Comparison of “Ca. Nitrosocaldales/Nitrosocaldaceae” with other AOA revealed 526 specific orthogroups. This included genes related to thermal adaptation (cyclic 2,3-diphosphoglycerate, and S-adenosylmethionine decarboxylase), indicating their importance for life at high temperature. In addition, these MAGs acquired genes from members from archaea (Crenarchaeota) and bacteria (Firmicutes), mainly involved in metabolism and stress responses, which might play a role to allow this group to adapt to thermal habitats.

A multi-omic screening approach for the discovery of thermoactive glycoside hydrolases.

Next-generation sequencing and computational biology have facilitated the implementation of new combinatorial screening approaches to discover novel enzymes of biotechnological interest. In this study, we describe the successful establishment of a multi-omic approach for the identification of thermostable hydrolase-encoding genes by determination of gene expression levels. We applied this combinatorial approach using an anaerobic enrichment culture from an Azorean hot spring sample grown on green coffee beans as recalcitrant substrate. An in-depth analysis of the microbial community resulted in microorganisms capable of metabolizing the selected substrate, such as the genera Caloramator,Dictyoglomus and Thermoanaerobacter as active and abundant microorganisms. To discover glycoside hydrolases, 90,342 annotated genes were screened for specific reaction types. A total number of 106 genes encoding cellulases (EC 3.2.1.4), beta-glucosidases (EC 3.2.1.21) and endo-1,4-beta-mannosidases (EC 3.2.1.78) were selected. Mapping of RNA-Seq reads to the related metagenome led to expression levels for each gene. Amongst those, 14 genes, encoding glycoside hydrolases, showed highest expression values, and were used for further cloning. Four proteins were biochemically characterized and were identified as thermoactive glycoside hydrolases with a broad substrate range. This work demonstrated that a combinatory omic approach is a suitable strategy identifying unique thermoactive enzymes from environmental samples.

Effect of pH on Different Strains of Phytase Producing Bacteria from Malaysia’s Hot Spring

In the recent research, the optimisation of culture condition for phytase production rarely done for Acetinobacter baumanii. The optimisation of the phytase production from the bacterial strains largely contributed by Bacillus sp. The study on the phytase originated from hot spring are limited and the species that identified from the hot spring samples are not in the same species from the previous study and mainly the species isolated from Bacillus sp. In this study, four potential strains of bacteria producing phytase isolated from hot spring in several regions in Malaysia. For enrichment of the bacterial, Nutrient Agar was used, meanwhile for batch culture optimisation, the bacteria producing phytase grown in modified liquid Phytase Screening Media with soy extract as agro residual substrate as a replacement for sodium phytate, the chemical substrate. The bacteria were screened for their ability to produce clear zone in solid PSM with sodium phytate as substrate. Optimisation of media through its physical factor that is pH of the media carried out using shake flask scale in laboratory. The growth of the bacterial strains and phytase activity measured quantitatively through the two different pH of media at pH 5.5 and pH 7. The analysis of colony-forming unit and pH determination after fermentation was carried out in this study. From the study, bacterial strain L3 from Labis, Johor has the highest phytase activity in the two parameters studied where the inorganic phosphate released at pH 5.5 (0.21953 U/mL) and pH 7 (0.2047 U/mL). Optimisation carried out through manipulating the culture condition that is pH of the media to determine at which condition has the highest phytase production. Several effects on enzyme activity caused by culture conditions identified. The optimisation of the fermentation medium able to contribute to the less cost production of the industrial enzyme as phytase has potential production for feed additives for poultry feeding. In the future research, molecular identification of the bacterial strains from the better-quality bacteria producing phytase grown in optimised culture media to investigate the molecular identity of the bacterial.