Phenotypic characterization for bioremediation suitability of isolates from Southern Tunisian tannery effluent.

Effluents from the leather tanning industry contain diverse pollutants, including hazardous heavy metals, posing threats to public health and the surrounding environment. Indigenous bacterial isolates can represent an eco-friendly approach for tannery wastewater treatment; however, phenotypic characterization is necessary to determine whether these strains are suitable for bioremediation. In the present study, we analyzed seven new Enterococcus faecium strains and two new Bacillus subtillis strains isolated from effluents from the Southern Tunisian Tannery (ESTT). We evaluated phenotypic features beneficial for bioremediation, including biofilm formation, hydrophobicity, and exoenzyme activities. Additionally, we examined characteristics naturally occurring in environmental bacteria but less desirable in strains selected for bioremediation, such as antibiotic resistances and pathogenicity indicators. The observed phenotypes were then compared with whole-genome analysis. We observed biofilm production in two slime-producing bacteria, B. licheniformis RLT6, and E. faecium RLT8. Hydrophobicity of E. faecium strains RLT1, RLT5, RLT8, and RLT9, as well as B. licheniformis RLT6 correlated positively with increasing ESTT concentration. Exoenzyme activities were detected in E. faecium strains RLT2, RLT4, and RLT7, as well as B. licheniformis RLT6. As anticipated, all strains exhibited common resistances to antibiotics and hemolysis, which are widespread in nature and do not hinder their application for bioremediation. Importantly, none of the strains exhibited the pathogenic hypermucoviscosity phenotype. To the best of our knowledge, this is the first report consolidating all these phenotypic characteristics concurrently, providing a complete overview of strains suitability for bioremediation. IMPORTANCE: The study evaluates the bioremediation potential of seven Enterococcus faecium strains and two Bacillus subtillis strains isolated from the effluents from the Southern Tunisian tannery (ESTT), which pose threats to public health and environmental integrity. The analysis primarily examines the phenotypic traits crucial to bioremediation, including biofilm formation, hydrophobicity, and exoenzyme activities, as well as characteristics naturally occurring in environmental bacteria related to heavy metal resistance, such as antibiotic resistances. Several strains were found to have high bioremediation potential and exhibit only antibiotic resistances commonly found in nature, ensuring their application for bioremediation remains uncompromised. The results of the exhaustive phenotypic analysis are contrasted with the whole genome sequences of the nine strains, underscoring the appropriateness of these bacterial strains for eco-friendly interventions in tannery wastewater treatment.

Biosorption of heavy metals from phosphate-processing effluent by Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads / Biosorción de metales pesados a partir de efluentes de procesamiento de fosfatos por Serratia rubidaea NCTC12971 inmovilizados en cuentas de Ca-alginato

In this study, the biosorption ability of various potentially toxic elements from phosphate-processing effluent (PPE) using the indigenous bacterium Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads was investigated. The experimental data analyzed by the Langmuir isotherm revealed that the optimum dose of 2 g·100 ml−1 of immobilized S. rubidaea NCTC12971 at pH 7 and a contact time of 48 h allowed the removal of 92.07%, 98.05%, 95.57%, and 88.39% of lead (Pb (II)), cadmium (Cd (II)), copper (Cu (II)), and zinc (Zn (II)), respectively. Moreover, under the Langmuir isotherm, the maximum single-layer adsorption capacity (qmax) of the biosorbent was estimated to 32.14 mg g−1, 45.87 mg g−1, 0.06 mg g−1, and 3.01 mg g−1 for Pb (II), Cd (II), Cu (II), and Zn (II), respectively, under the stated conditions. Alternatively, the regeneration and reuse of the Ca-alginate beads was evaluated. Indeed, after four consecutive adsorption–desorption cycles, there was no significant loss in the biosorption capacity. The effectiveness of the bacterial biosorption as treatment process was evaluated by assessing the phytotoxicity of the treated effluent (TE) on Medicago sativa and Lactuca sativa seed germination and their root elongation. Results exhibited a significant toxicity removal expressed by a notable increase in the germination indices (GI), which reach 80% and 70%, respectively, for Medicago sativa and Lactuca sativa compared to the GI values of 46.6% and 16.6% of the same species in presence of the untreated effluent (PPE).(AU)

Biosorption of heavy metals from phosphate-processing effluent by Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads.

In this study, the biosorption ability of various potentially toxic elements from phosphate-processing effluent (PPE) using the indigenous bacterium Serratia rubidaea NCTC12971 immobilized in Ca-alginate beads was investigated. The experimental data analyzed by the Langmuir isotherm revealed that the optimum dose of 2 g·100 ml-1 of immobilized S. rubidaea NCTC12971 at pH 7 and a contact time of 48 h allowed the removal of 92.07%, 98.05%, 95.57%, and 88.39% of lead (Pb (II)), cadmium (Cd (II)), copper (Cu (II)), and zinc (Zn (II)), respectively. Moreover, under the Langmuir isotherm, the maximum single-layer adsorption capacity (qmax) of the biosorbent was estimated to 32.14 mg g-1, 45.87 mg g-1, 0.06 mg g-1, and 3.01 mg g-1 for Pb (II), Cd (II), Cu (II), and Zn (II), respectively, under the stated conditions. Alternatively, the regeneration and reuse of the Ca-alginate beads was evaluated. Indeed, after four consecutive adsorption-desorption cycles, there was no significant loss in the biosorption capacity. The effectiveness of the bacterial biosorption as treatment process was evaluated by assessing the phytotoxicity of the treated effluent (TE) on Medicago sativa and Lactuca sativa seed germination and their root elongation. Results exhibited a significant toxicity removal expressed by a notable increase in the germination indices (GI), which reach 80% and 70%, respectively, for Medicago sativa and Lactuca sativa compared to the GI values of 46.6% and 16.6% of the same species in presence of the untreated effluent (PPE).

Study of the phytotoxic potential of olive mill wastewaters on a leguminous plant 'Vicia faba L.'

We investigated the study of the phytotoxic potential of olive mill wastewaters (OMW) on the germination and growth responses of the faba bean 'Vicia faba L.' leguminous plant cultivated under an arid Mediterranean climate. The results showed that the raw OMW blocked the germination of the seeds tested, while OMW treated soil extracts stimulate the seeds' germination rate. The monitoring of the faba bean growth parameters in the different OMW treated soils showed that the optimal growth of the faba bean plants has been recorded for the soil amended with 25 m3 ha-1 of OMW. Although the 50 m3 ha-1 dose is the most suitable for the soil studied, our results showed that the 25 m3 ha-1 dose is the most suitable for the vegetative development, as well as for the productivity of the plant tested 'Vicia faba L.'.

Influence of biowaste compost amendment on soil organic carbon storage under arid climate.

Organic matter amendments have been proposed as a means to enhance soil carbon stocks on degraded soils, particularly under arid climate. Soil organic carbon (SOC) plays a critical role in terrestrial carbon cycling and is central to preserving soil quality. The effects of biowaste compost (BWC) on soil carbon storage were investigated. In addition, changes in soil organic matter (SOM) and even soil organic carbon (SOC) in BWC-amended soils following different applications were studied. The added BWC quantities were as followed: BWC/soil (weight/weight (w/w) respectively: 1/8, 1/4, and 1/2). The different BWC-amended soils were assessed during 180 days under arid ambient conditions and in comparison with control soil. Results showed a significant increase in SOM and SOC with relation to BWC quantities applied. This increase was relatively clear up to 120 days, after which decrease in SOM and SOC levels were observed. Furthermore, results showed improved microbiological activities of the amended soils in comparison with the control soil. This was reflected by the increase of the amended soils' respirometric activities as cumulative carbon dioxide carbon (C-CO2) as function of incubation time and also in terms of specific respiration expressed as C-CO2/SOC ratios. Implications: Mediterranean soils under arid climate such as Tunisian soils are poor in organic matter content. Biowastes are potential source for soil fertilization. Composting process is the best method for the stabilization of organic matter of diverse origins. The biowaste compost amendment improves the soil organic carbon storage and enhances the soil microbial activity.

Study of Heavy Metal Accumulation and Residual Toxicity in Soil Saturated with Phosphate Processing Wastewater.

The effects of phosphate processing wastewater (PPWW) on heavy metal accumulation in a Mediterranean soil (Tunisia, North Africa) were investigated. Moreover, the residual toxicities of PPWW-irrigated soils extracts were assessed. Results showed that heavy metal accumulation was significantly higher in PPWW-irrigated soil extracts than in control soil. The heavy metal accumulation increased over time in treated soil samples and their average values followed the following order: Iron (Fe 252.72 mg l-1) > Zinc (Zn 152.95 mg l-1) > Lead (Pb 128.35 mg l-1) > Copper (Cu 116.82 mg l-1) > Cadmium (Cd 58.03 mg l-1). The residual microtoxicity and phytotoxicity of the various treated soil samples extracts were evaluated by monitoring the bioluminescence inhibition (BI %) of Vibrio ficheri and the measurement of the germination indexes (GI %) of Lepidium sativum and Medicago sativa seeds. The results showed an important increase of residual toxicities of PPWW-treated soil extracts over time.

Assessment of toxicity of the untreated and treated olive mill wastewaters and soil irrigated by using microbiotests.

Hazard assessments based on two measures of toxicity were conducted for the untreated olive mill wastewaters (U), untreated olive mill wastewaters organic extract (UOE), treated olive mill wastewaters (T), treated olive mill wastewaters organic extract (TOE) and extracts of soils ferti-irrigated with untreated (SU) and with treated olive mill wastewaters (ST). The measures of toxicity were achieved by the determination of the bioluminescence inhibition percent (I(B)%) of Vibrio fischeri and by the growth inhibition (GI) of Bacillus megaterium, Pseudomonas fluorescens and Escherichia coli. A bioluminescence inhibition of V. fischeri of 100%, 100%, 65%, 47%, 46% and 30% were obtained with U, UOE, T, TOE, SU and ST respectively. Indeed, even diluted 24 times, a significant bioluminescence inhibition of 96% was obtained by U. However, only 30% bioluminescence inhibition was obtained by 24 times diluted T. Whereas, 24 times diluted, SU and ST did not show a bioluminescence inhibition (3% and 1%, respectively). The GI of B. megaterium, P. fluorescens and E. coli were, respectively, 93%, 72% and 100% by U; 100%, 80% and 100% by UOE; 70%, 60% and 89% by T; 63%, 54% and 68% by TOE; 39%, 27% and 43% by SU and 23%, 0% and 34% by ST. The incubation of U or T in the soil during four months reduced their toxicity by 54% and 35%, respectively. As it was expected, the most resistant bacterium to OMW toxicity is P. fluorescens then B. megaterium and E. coli. V. fischeri remained the most sensitive strain to the toxicity of this sewage what proves again its utilisation as standard of measure of the toxicity.

Polyphenols dynamics and phytotoxicity in a soil amended by olive mill wastewaters.

The effects of unprocessed olive mill wastewaters (OMW) on soil characteristics were investigated. Phenolic compounds levels in the treated soil were compared to those of a control soil profile. Results showed that OMW infiltration caused a modification of soil physicochemical characteristics. Phenolic compounds were detected at a depth of 1.2m four months after the last application of OMW. A moderate phytotoxic residual phenolic fraction (F) was extracted from the superficial soil layer 1 year after the OMW application. This residual F had a phytotoxic potential comparable to that of 25-fold diluted OMW.

Changes in microbial and soil properties following amendment with treated and untreated olive mill wastewater.

We investigated the effect of untreated and biologically treated olive mill wastewater (OMW) spreading on the soil characteristics and the microbial communities. The water holding capacity, the salinity and the content of total organic carbon, humus, total nitrogen, phosphate and potassium increased when the spread amounts of the treated or untreated OMW increased. The OMW treated soil exhibited significantly higher respiration compared to the control soil. However, the C-CO2/C(tot) ratio decreased from 1.7 in the control soil to 0.5 in the soil amended with 100 m3 ha(-1) of untreated OMW. However, it slightly decreased to 1.15 in the soil amended with 400 m3 ha(-1) of treated OMW. The treated OMW increased the total mesophylic number while the number of fungi and nitrifiers decreased. Actinomycetes and spore-forming bacteria were neither sensitive to treated nor to untreated OMW. The total coliforms increased with higher doses of treated and untreated OMW. A toxic effect of the untreated OMW appeared from 100 m3 ha(-1). This toxicity was more significant with 200 m3 ha(-1), where microflora of total mesophilic, yeasts and moulds, actinomycetes, and nitrifiers were seriously inhibited except for total coliforms and spore-forming bacteria.