Soil extracts are useful nutrients to enhance the growth of microalgae. Therefore, the present study attempts for the use of virgin soils from Peninsular Malaysia as growth enhancer. Soils collected from Raja Musa Forest Reserve (RMFR) and Ayer Hitam Forest Reserve (AHFR) were treated using different extraction methods. The total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and dissolved organic carbon (DOC) concentrations in the autoclave methods were relatively higher than natural extraction with up to 132.0 mg N/L, 10.7 mg P/L, and 2629 mg C/L, respectively for RMFR. The results of TDN, TDP, and DOC suggested that the best extraction methods are autoclaved at 121 °C twice with increasing 87%, 84%, and 95%, respectively. Chlorella vulgaris TRG 4C dominated the growth at 121 °C twice extraction method in the RMRF and AHRF samples, with increasing 54.3% and 14%, respectively. The specific growth rate (µ) of both microalgae were relatively higher, 0.23 d⁻¹ in the Ayer Hitam Soil. This extract served well as a microalgal growth promoter, reducing the cost and the needs for synthetic medium. Mass production of microalgae as aquatic feed will be attempted eventually. The high recovery rate of nutrients has a huge potential to serve as a growth promoter for microalgae.

Soil ecosystems are home to a diverse range of microorganisms, but they are only partially understood because no single-cell sequencing or whole-community sequencing provides a complete picture of these complex communities. Using one of such metagenomics approaches, we succeeded in monitoring the microbial diversity and stress-response gene in the soil samples. This study aims to test whether known differences in taxonomic diversity and composition are reflected in functional gene profiles by implementing whole gene sequencing (WGS) metagenomic analysis of geographically dispersed soils from two distinct pristine forests. The study was commenced by sequencing three rainforest soil samples and three peat swamp soil samples. Soil richness effects were assessed by exploring the changes in specific functional gene abundances to elucidate physiological constraints acting on different soil systems and identify variance in functional pathways relevant to soil biogeochemical cycling. Proteobacteria shows abundances of microbial diversity for 52.15% in Royal Belum Reserved Forest and 48.28% in Raja Musa; 177 out of 1,391,841 and 449 out of 3,586,577 protein coding represent acidic stress-response genes for Royal Belum and Raja Musa, respectively. Raja Musa indicates pH 2.5, which is extremely acidic. The analysis of the taxonomic community showed that Royal Belum soils are dominated by bacteria (98% in Sungai Kooi (SK), 98% in Sungai Papan (SP), and 98% in Sungai Ruok (SR), Archaea (0.9% in SK, 0.9% in SP, and 1% in SR), and the remaining were classed under Eukaryota and viruses. Likewise, the soils of Raja Muda Musa are also dominated by bacteria (95% in Raja Musa 1 (RM1), 98% in Raja Musa 2 (RM2), and 96% in Raja Musa 3 (RM3)), followed by Archaea (4% in RM1, 1% in RM2, and 3% in RM3), and the remaining were classed under Eukaryota and viruses. This study revealed that RBFR (Royal Belum Foresr Reserve) and RMFR (Raja Musa Forest Reserve) metagenomes contained abundant stress-related genes assigned to various stress-response pathways, many of which did not show any difference among samples from both sites. Our findings indicate that the structure and functional potential of the microbial community will be altered by future environmental potential as the first glimpse of both the taxonomic and functional composition of soil microbial communities.

The assessment of water-extractable organic matter using an autoclave can provide useful information on physical, chemical, and biological changes within the soil. The present study used virgin forest soils from Chini Forest Reserve, Langkawi Island, and Kenyir Forest Reserve (Malaysia), extracted using different extraction methods. The dissolved organic carbon (DOC), total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and ammonium-nitrate content were higher in the autoclave treatments, up to 3.0, 1.3, 1.2, and 1.4 times more than by natural extraction (extracted for 24 h at room temperature). Overall, the highest extractable DOC, TDN, TDP, ammonium and nitrate could be seen under autoclaved conditions 121 °C 2×, up to 146.74 mg C/L, 8.97 mg N/L, 0.23 mg P/L, 5.43 mg N mg/L and 3.47 N mg/L, respectively. The soil extracts became slightly acidic with a higher temperature and longer duration. Similar trends were observed in the humic and nonhumic substances, where different types of soil extract treatments influenced the concentrations of the fractions. Different soil extraction methods can provide further details, thus widening the application of soil extracts, especially in microbes.

Microalgae are widely utilized in commercial industries. The addition of a modified artificial medium (soil extract) could enhance their growth. Soil extract collected from the Raja Musa peat swamp and mineral soil from the Ayer Hitam Forest Reserve (AHFR), Selangor, Malaysia, were treated using various extraction methods. Carteria radiosa PHG2-A01, Neochloris conjuncta, and Nephrochlamys subsolitaria were grown in microplates at 25 °C, light intensity 33.75 µmol photons m⁻²s⁻¹ for 9 days. N. conjuncta dominated the growth in 121 °C twice extraction method AFHR samples, with 47.17% increment. The highest concentrations of ammonia and nitrate were detected in the medium with soil extract treated with 121 °C twice extraction method, yielding the concentrations of 2 mg NL⁻¹ and 35 mg NL⁻¹ for ammonia and nitrate of RM soil and 2 mg NL⁻¹ and 2.85 mg NL⁻¹ for the AH soil. These extracts are proved successful as a microalgal growth stimulant, increasing revenue and the need for enriched medium. The high rate of nutrient recovery has the potential to serve as a growth promoter for microalgae.

– Soil pH affects the microbial community's composition and diversity, although research on microorganisms in preserved peat swamp forests is rare. In terms of soil acidity, peat swamp forest has been classified as one of the most nutrient-deficient areas of the planet. To have a more comprehensive understanding of the ecology and function of these microbial communities, profiling of peat swamp soil microbial community and structure was conducted with Raja Musa Peat Swamp Forest Reserved (RMFR) in Selangor for its pristine soil that refers to its stability of the microbial community and soil compositions, while focusing on pH response against the microbial community. This study was conducted by using Whole Genome Sequences analysis complemented with Next Generation Sequencing (NGS) and whole shotgun metagenomic. The focus was to clarify whether the naturally occurring pH variations, specifically acid stress and osmotic stress in stress response, affected the soil microbial functional biodiversity and composition. Details analysis from MG-RAST has shown that bacteria has dominated the community with 98 percent abundance while eucaryote with 1 percent and archaea 1 percent. At the phylum level, the four highest abundances recorded which Proteobacteria hits with 51 percent abundances, Actinobacteria 18 percent, Acidobacteria 9 percent, and firmicutes 7 percent. At the genus level, the five highest abundances have been identified: Burkholderia has recorded 6 percent abundances, Candidatus Solibacter 4 percent of abundances, Bradyrhizobium has 4 percent abundances, Streptomyces 4 percent of abundances, and Rhodopseudomonas has 3 percent of abundances,. Amino acid metabolism analysis has identified a significant percentage of valine, leucine, and isoleucine involved with stress response genes that contribute to acid stress and osmotic stress with the microbial genus of the highest abundances has involved where Burkholderia has 10 percent abundances, Bradyrhizobium 10 percent abundances, Rhodopseudomonas 10 percent of abundances, Mycobacterium 8 percent abundances, and Frankia 7 percent abundances.

The microalgae-based industries are trending upwards, particularly as the feed ingredient for aquaculture. Therefore, a sustainable and reasonably priced source of nutrients to support the mass cultivation of microalgae is in great demand. The present study explored the feasibility of using extracts from virgin soil as natural growth-promoting nutrients for the cultivation of Nannochloropsis oculata, Nannochloropsis oceanica, and Chlorella sorokiniana. The extracts were obtained from Bera Lake Forest using five different treatment methods. The greatest retrieval of dissolved organic carbon, total dissolved nitrogen, and total dissolved phosphorus were observed with the autoclave treatment method at 121 °C twice, yielding a respective concentration of 336.56 mg/L, 13.40 mg/L, and 0.14 mg/L, respectively. The highest growth was recorded with Nannochloropsis oculata resulting in an optical density of 0.488 ± 0.009 (×10³ cell mL⁻¹), exhibiting 43% and 44% enhanced growth in comparison to Nannochloropsis oceanica and Chlorella sorokiniana, respectively. The specific growth rate (0.114 ^a ± 0.007 d⁻¹) was the highest for Nannochloropsis oculata when the 24 h-extraction method was used, whereas the utilization of the autoclave 121 °C twice treatment method contributed to the highest specific growth of Nannochloropsis ocenica (0.069 ^a ± 0.003 d⁻¹) and Chlorella sorokiniana (0.080 ^a ± 0.001 d⁻¹). Collectively, these findings suggested that the addition of soil extracts which is sustainable and inexpensive promoted the growth of microalgae compared to the control system. A further study investigating the optimum culture conditions for enhanced microalgae growth will be carried out for the mass production of microalgae biomass.