The bio-flocculants used in this study were synthesised by the Mannich reaction, which includes three reagents: a substrate (tannin extracts of Acacia, Quebracho, and Castanea), formaldehyde, and an amine derivative (ethanolamine, diethanolamine, ammonium chloride). Nine natural flocculants were prepared by combining extracts and amines; these products were evaluated in three different wastewater samples in two experimental phases. In phase I, five physicochemical parameters were analysed. From the data obtained, a multivariate, completely randomised design (CRD-Manava) was used, with a factorial arrangement and mean plots. In phase II, the three bio-flocculants with the most statistically significant responses and their mixtures were examined, evaluating 14 biological and physicochemical parameters. Statistical analysis was guided in this phase by CRD blocks, finding a significant removal in the physicochemical parameters analysed in the different types of wastewater and obtaining removal rates between 50 and 90%, depending on the parameter. At the end of both phases, the bio-flocculants acacia-ammonium chloride and quebracho-diethanolamine were the most efficient in the removal of turbidity (34–99%), true colour (93–100%) and total solids (12–99%). In addition, the natural flocculants showed low mutagenicity index (MI: 0.33–0.93) compared to aluminium sulphate (MI: 4.87–8.81).

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Berberis goudotii is an endemic Colombian plant found in the paramo ecosystem. It has been used in food preparation and as a medicinal plant for diverse treatments. Additionally, it is used as a mouthwash to strengthen the gums and combat throat irritations and periodontitis. The present research evaluated Berberis goudotii aerial parts extract and fractions antimicrobial activities. Ultrasonic-assisted extraction was used to attain total ethanol-water extract. Solid-liquid fractionation was used to obtain hexane fraction. The residue was dispersed in water and liquid-liquid fractionation was carried-out to acquire dichloromethane, butanol and water fractions. Preliminary phytochemical analysis was performed on total extract and phenol, polyphenol, flavonoid, and proanthocyanidin, while tannin content was quantified. Antimicrobial activity assessment was performed by agar diffusion method using disks and wells employing Ceftazidime as a positive control against Streptococcus mutans, Streptococcus sobrinus, Lactobacillus acidophilus, Lactobacillus casei, Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. Antimicrobial activity was determined as relative percentage inhibition (RPI), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Phenols (92.5 ± 7.7 mg GA/10 g), polyphenols (87.7 ± 8.1 mg PG/10 g) and tannins (44.1 ± 4.3 mg PG/10 g) were among the highest secondary metabolites observed. Total extract presented an MBC of 1.0 µg/µL against cariogenic bacteria (Streptococcus mutans and Streptococcus sobrinus) and 0.12 µg/µL against bacteria associated with periodontal disease (Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum). Butanol and hexane fractions showed antiperiodontal activity with MBC of 0.12 and 1.0 µg/µL, respectively. In conclusion, Berberis goudotii total extract demonstrated antimicrobial activity against cariogenic and periodontal microorganisms, on the other hand, hexane and butanol fractions displayed antiperiodontal activity.