Folate, an essential nutrient crucial for DNA synthesis, cell division, and fetal neural tubedevelopment, remains a global health concern due to deficiencies in certain populations. Toaddress this issue, we investigated the synergistic interaction between Spirulina, a nutrient-rich microalga, and two strains of folic acid-producing bacteria, Bacillus subtilis-1 and Bacillus subtilis-2, to enhance folate production. Spirulinahas high nutritional content, combined with the folate production capabilities of the selected bacteria, offered a promising opportunity for sustainable folate synthesis. In this study, Spirulinaand the two strains of Bacillus subtilis were cultured separately to optimize growth conditions for each organism. Co-culture experiments were then conducted, combining Spirulina with Bacillus subtilis-1 and also Bacillus subtilis-2, to investigate their collective potential for folate production. The specific growth rates of both Spirulinaand the bacteria were measured individually and in combination using spectrophotometric methods, and their dry weights were determined to assess biomass productivity. Folate quantification in the microalgal-bacterial cultures was performed using a spectrophotometric analysis based on the phosphate buffer extraction method. This method facilitated the measurement of folate content investigated the impact of the symbiotic relationship between Spirulina and bacteria, particularly in terms of enhancing vitamin B12 acquisition and its impact on folate synthesis. Our results revealed a synergistic enhancement in folate production at the exponential growth phase of Bacillus subtilis-1 and Bacillus subtilis-2 co-culture. The synergistic relationship between Spirulina and Bacillus subtilis-1, as well as Bacillus subtilis-2, is manifested in elevated folate synthesis, highlighting the significant impact of microbial cooperation on nutrient production. Additionally, we observed fluctuations in folate production at the stationary phase, highlighting the subtle equilibrium achieved through symbiotic interactions. These findings shed light on the potential of harnessing the synergistic potential of microorganisms for sustainable folate synthesis. This research advances co-culturing methods to optimize folate production, tackling global folate deficiency challenges and fostering innovative, sustainable nutritional solutions.

This study aimed to assess the impact of individual as well as combined application of Lysinibacillus macroides and Bacillus safensis in phytoremediation potential of Brassica napus grown in soil contaminated by industrial effluents. In response to five metals; copper, chromium, nickel, lead, and cadmium, results revealed that germination percentage, fresh and dry weights, and photosynthetic pigments of B. napus decreased under contaminated soil. On the other hand, electrolyte leakage due to cellular injury, metabolites (proline and glycine betaine), antioxidant enzymes (superoxide dismutase, catalase, peroxidase, ascorbate peroxidase), accumulation of hydrogen peroxide and metals in plant’s roots, shoots and leaves increased. Inoculation significantly reduced these effects as proved by the enhancement of germination percentage, fresh and dry biomass, and photosynthetic pigments. Simultaneously, the antioxidant enzymes, metabolites contents (proline and glycine betaine) and metal concentrations in plant’s roots, shoots and leaves decreased. Combined application of both Bacilli strains was found more effective as compared to individual inoculation. It was concluded that metal resistant Bacillus species in combination had growth effects on B. napus and enhanced its phytoremediation efficiency in contaminated soil. Novelty statement Brassica napus; a hyper-accumulator of metals, loses phytoremediation potential with the passage of growth. Two Bacillus species (Lysinibacillus macroides and Bacillus safensis) having known bioremediation abilities were employed individually as well as in combination under metals contaminated soil to increase phytoremediation efficiency of B. napus. The metals containing soil used is a unique aspect in this study because selected soil, contaminated by industrial effluents, has not been evaluated or reported earlier. Combined application of Bacilli improved phytoremediation potential of B. napus more as compared to application of individual Bacillus strain which is yet another unique aspect of this investigation.

This pioneering study uniquely identifies and characterizes the presence of algal species, mainly Spirulina, from water reservoirs in the Kohat district of Pakistan, a previously unstudied region for microalgae dynamics. Conducted over one year, from July 2022 to June 2023, the study examined 156 samples from 26 freshwater bodies in Kohat. Only one sample from Usterzai (pH 8.6 ± 0.08, TDSs 313 ± 0.81 mg/L, DO 4.50 ± 0.05 mg/L, EC 540 ± 0.81 µS/cm) contained Spirulina sp., highlighting its rarity and specific environmental preferences. The other 155 samples, with varying parameters, contained different microalgae. Microscopic analysis further confirmed the presence of Spirulina in only one sample. The morphological and molecular analyses of the isolated Spirulina culture showed variability within the population, with phylogenetic analysis illuminating closer relationship with Arthrospira platensis. While multivariate analyses identified key environmental parameters influencing algal species distribution, the selective presence of Spirulina was found less relevant, which requires further investigation in terms of nutrient availability, microbial interactions, or subtle variations in water chemistry for ecological preferences and adaptations.