Microalgae produced during phycoremediation of swine wastewater contains effective bacteriostatic compounds against antibiotic-resistant bacteria.

Studies on the antimicrobial effects of microalgae extracts are commonly reported using algae biomass grown in sterile synthetic mineral medium and controlled laboratory conditions. However, variations in environmental conditions and culture medium composition are known to alter microalgae biochemical structure possibly affecting the type and concentrations of bioactive compounds with antimicrobial properties. In this work, solvent extracts of the microalgae Chlorella spp. were tested for antimicrobial effects against gram-positive and multidrug resistant pathogenic bacteria Staphylococcus hyicus, Enterococcus faecalis and Streptococcus suis. Microalgae was cultivated at field scale open pond reactor using raw swine wastewater as growth substrate. Dichloromethane or methanol were used to obtain the microalgae extracts. Characterization of the extracts by ultra-high performance liquid chromatography-quadrupole mass spectrometry revealed the presence of 23 phytochemicals with recognized antimicrobial properties. Bacteriostatic activity was observed in plating assays by formation of inhibition zones ranging from 7 to 18 mm in diameter. Only dichloromethane extracts were inhibitory to all three model bacteria. The minimum inhibitory concentration assessed for dichloromethane extracts were 0.5 mg mL-1 for Staphylococcus hyicus and Enterococcus faecalis and 0.2 mg mL-1 for Streptococcus suis. Bactericidal effects were not observed using solvent-extracts at 2 or 5 mg L-1. To the best of authors knowledge, this is the first report on the antimicrobial effects of Chlorella spp. extracts against Staphylococcus hyicus and Streptococcus suis. Overall, Chlorella spp. grown on swine wastewater contains several phytochemicals that could be further explored for the treatment of infections caused by antibiotic-resistant bacteria pathogens.

Crotalaria spectabilis as a source of pyrrolizidine alkaloids and phenolic compounds: HPLC-MS/MS dereplication and monocrotaline quantification of seed and leaf extracts.

INTRODUCTION: Crotalaria spectabilis is an important species used as a pre-plant cover for soybean crops to control the proliferation of endoparasitic nematodes. Species from the Crotalaria genus are known for presenting pyrrolizidine alkaloids (PAs) in their composition, however, C. spectabilis is still considered chemically under-explored. OBJECTIVE: The goal of this manuscript is the development and validation of a method for PAs and flavonoids identification and quantification of C. spectabilis seeds and leaves, a toxic plant used for nematode proliferation control in soil, especially in soybean crops. MATERIALS AND METHODS: Seeds and leaves extracts were analysed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) for the identification of the compounds. RESULTS: PAs and phenolic compounds could be identified in both samples based on the MS/MS fragmentation pattern. Molecular formulas of the annotated compounds were confirmed by ultra-high-performace liquid chromatography-quadrupole time-of-flight (UHPLC-QToF), and monocrotaline could also be confirmed by standard comparison. The quantification of monocrotaline was performed by HPLC-MS/MS, resulting in 123 times higher monocrotaline content in seeds than in the leaves, which could explain its efficiency in combating nematode proliferation in soil. CONCLUSION: This was the first report of phenolic compounds in C. spectabilis. The current study highlights the importance of C. spectabilis for nematode control due to the presence of toxic PAs, and the employment of analytical techniques for identification and quantification of compounds present in the extracts.

Unraveling Asian Soybean Rust metabolomics using mass spectrometry and Molecular Networking approach.

Asian Soybean Rust (ASR), caused by the biotrophic fungus Phakopsora pachyrhizi, is a devastating disease with an estimated crop yield loss of up to 90%. Yet, there is a nerf of information on the metabolic response of soybean plants to the pathogen Untargeted metabolomics and Global Natural Products Social Molecular Networking platform approach was used to explore soybean metabolome modulation to P. pachyrhizi infection. Soybean plants susceptible to ASR was inoculated with P. pachyrhizi spore suspension and non-inoculated plants were used as controls. Leaves from both groups were collected 14 days post-inoculation and extracted using different extractor solvent mixtures. The extracts were analyzed on an ultra-high performance liquid chromatography system coupled to high-definition electrospray ionization-mass spectrometry. There was a significant production of defense secondary metabolites (phenylpropanoids, terpenoids and flavonoids) when P. pachyrhizi infected soybean plants, such as putatively identified liquiritigenin, coumestrol, formononetin, pisatin, medicarpin, biochanin A, glyoceollidin I, glyoceollidin II, glyoceollin I, glyoceolidin II, glyoceolidin III, glyoceolidin IV, glyoceolidin VI. Primary metabolites (amino acids, peptides and lipids) also were putatively identified. This is the first report using untargeted metabolomics and GNPS-Molecular Networking approach to explore ASR in soybean plants. Our data provide insights into the potential role of some metabolites in the plant resistance to ASR, which could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.