Comparative study between Salkowski reagent and chromatographic method for auxins quantification from bacterial production.

Introduction: The Salkowski reagent method is a colorimetric technique used to determine auxin production, specifically as indole-3-acetic acid (IAA). It was developed to determine indoles rapidly; however, it does not follow Beer's law at high concentrations of IAA. Thus, there could be an overestimation of IAA with the Salkowski technique due to the detection of other indole compounds. Methods: This study aims to compare the Salkowski colorimetric method versus a chromatographic method to evidence the imprecision or overestimation obtained when auxins, such as indole-acetic acid (IAA), are determined as traits from promoting growth plant bacteria (PGPB), using ten different strains from three different isolation sources. The analysis used the same bacterial culture to compare the Salkowski colorimetric and chromatographic results. Each bacterium was cultivated in the modified TSA without or with tryptophan for 96 h. The same supernatant culture was used in both methods: Salkowski reagent and ultra-performance liquid chromatography coupled with a Mass Spectrometer (LC-MS/MS). Results: The first method indicated 5.4 to 27.4 mg L-1 without tryptophan in ten evaluated strains. When tryptophan was used as an inductor of auxin production, an increase was observed with an interval from 4.4 to 160 mg L-1. The principal auxin produced by all strains was IAA from that evaluated by the LC-MS/MS method, with significantly higher concentration with tryptophan addition than without. Strains belonging to the Kocuria genus were highlighted by high IAA production. The indole-3-propionic acid (IPA) was detected in all the bacterial cultures without tryptophan and only in K. turfanensis As05 with tryptophan, while it was not detected in other strains. In addition, indole-3-butyric acid (IBA) was detected at trace levels (13-16 µg L-1). Conclusions: The Salkowski reagent overestimates the IAA concentration with an interval of 41-1042 folds without tryptophan and 7-16330 folds with tryptophan as inductor. In future works, it will be necessary to determine IAA or other auxins using more suitable sensitive techniques and methodologies.

PGPB Consortium Formulation to Increase Fermentable Sugar in Agave tequilana Weber var. Blue: A Study in the Field.

Agave tequilana Weber var. Blue is used as the primary raw material in tequila production due to its fructans (inulin) content. This study evaluates the formulation of a plant-growth-promoting bacteria (PGPB) consortium (Pseudomonas sp. and Shimwellia sp.) to increase sugars in A. tequilana under field conditions. A total of three doses were tested: low (5 L ha-1), medium (10 L ha-1), and high (15 L ha-1), with a cellular density of 1 × 108 CFU mL-1 and one control treatment (without application). Total reducing sugars (TRS), inulin, sucrose, glucose, fructose, and plant growth were measured in agave plants aged 4-5 years at 0 (T0), 3 (T3), 6 (T6), and 12 (T12) months. Yield was recorded at T12. The TRS increased by 3%, and inulin by 5.3% in the high-dose treatment compared to the control at T12. Additionally, a low content of sucrose, glucose, and fructose (approximately 1%) was detected. At T12, the weight of agave heads increased by 31.2% in the medium dose and 22.3% in the high dose compared to the control. The high dose provided a higher inulin content. The A. tequilana plants were five years old and exhibited growth comparable to the standards for 6-7-year-old plants. This study demonstrates a sustainable strategy for tequila production, optimizing the use of natural resources and enhancing industry performance through increased sugar content and yield.

Use of agave bagasse for production of an organic fertilizer by pretreatment with Bjerkandera adusta and vermicomposting with Eisenia fetida.

Agave tequilana Weber is used in tequila and fructans production, with agave bagasse generated as a solid waste. The main use of bagasse is to produce compost in tequila factories with a long traditional composting that lasts 6-8 months. The aim of this study was to evaluate the degradation of agave bagasse by combining a pretreatment with fungi and vermicomposting. Experiments were carried out with fractionated or whole bagasse, sterilized or not, subjecting it to a pretreatment with Bjerkandera adusta alone or combined with native fungi, or only with native bagasse fungi (non-sterilized), for 45 days. This was followed by a vermicomposting with Eisenia fetida and sewage sludge, for another 45 days. Physicochemical parameters, lignocellulose degradation, stability and maturity changes were measured. The results indicated that up to 90% of the residual sugars in bagasse were eliminated after 30 days in all treatments. The highest degradation rate in pretreatment was observed in non-sterilized, fractionated bagasse with native fungi plus B. adusta (BNFns) (71% hemicellulose, 43% cellulose and 71% lignin) at 45 days. The highest total degradation rates after vermicomposting were in fractionated bagasse pre-treated with native fungi (94% hemicellulose, 86% cellulose and 91% lignin). However, the treatment BNFns showed better maturity and stability parameters compared to that reported for traditional composts. Thus, it seems that a process involving vermicomposting and pretreatment with B. adusta could reduce the degradation time of bagasse to 3 months, compared to the traditional composting process, which requires from 6 to 8 months.

Eisenia fetida increased removal of polycyclic aromatic hydrocarbons from soil.

The removal of phenanthrene, anthracene and benzo(a)pyrene added at three different concentrations was investigated with or without earthworms (Eisenia fetida) within 11 weeks. Average anthracene removal by the autochthonous micro-organisms was 23%, 77% for phenanthrene and 13% for benzo(a)pyrene, while it was 51% for anthracene, 47% for benzo(a)pyrene and 100% for phenanthrene in soil with earthworms. At 50 and 100mg phenanthrene kg(-1)E. fetida survival was 91% and 83%, but at 150 mg kg(-1) all died within 15 days. Survival of E. fetida in soil amended with anthracene < or = 1000 mg kg(-1) and benzo(a)pyrene < or = 150 mg kg(-1) was higher than 80% and without weight loss compared to the untreated soil. Only small amounts of PAHs were detected in the earthworms. It was concluded that E. fetida has the potential to remove large amounts of PAHs from soil, but more work is necessary to elucidate the mechanisms involved.