Spectroscopic and microscopic characterization of humic acids from composts made by co-composting of green waste, spent coffee and OMWW sludge.

Due to its important role in the formation of humic acids (HA), improving lignin degradation during composting has usually been considered a challenge. One practice that could stimulate the biodegradation of this recalcitrant molecule is inoculation with exogenous lignolytic fungal strains. Two composts (C1) and (C2) from piles (H1) and (H2) were evaluated. H1 was the control pile and H2 was inoculated at maturity with Trametes trogii, resulting in a 35% increase in lignin degradation rate compared to H1. The aim of this study was to show the main effects of this increase on the humification process in the co-composting of green waste, coffee grounds and olive mill wastewater sludge (OMWWs). Microstructure of HA1 and HA2 extracted from C1 and C2, respectively, was also investigated by scanning electron microscopy (SEM) and SEM coupled with energy-dispersive X-ray spectroscopy (X-EDS). The results showed that there were several similarities between the compost samples tested. These included the mineral content, the degree of polymerization (PD)> 1 and the compact and rigid surface of the extracted HA. However, C2 was characterized by a higher humic content (HC), degree of polymerization (PD), humification index (HI) and percentage of humic acids (PHA) than C1. Carbon-13 nuclear magnetic resonance (13C-NMR) and Fourier transmission-infrared spectroscopy (FTIR) analysis showed that aliphatic groups such as hydroxyls, alcohols and carboxyls were predominant in both composts. SEM analysis in conjunction with X-EDS analysis of HA2 showed a higher proportion of carbon and potassium (18 and 7.93%) than in HA1 (14 and 0.95%).

Activated sludge and UV-C254 for Sapovirus, Aichivirus, Astrovirus, and Adenovirus processing.

In this study, the detection rates of four enteric viruses, Human Astrovirus (HAstVs), Aichivirus (AiVs), Human Adenovirus (HAdVs), and Sapovirus (SaVs) are carried out to assess the virological quality of the treated wastewater. A total of 140 samples was collected from wastewater treatment plant WWTP of Tunis-City. Real-time RT-PCR and conventional RT-PCR results showed high frequencies of detection of the four enteric viruses investigated at the entry and exit of the biological activated sludge procedure and a significant reduction in viral titers after tertiary treatment with UV-C254 irradiation. These results revealed the ineffectiveness of the biological activated sludge treatment in removing viruses and the poor quality of the treated wastewater intended for recycling, agricultural reuse, and safe discharge into the natural environment. The UV-C254 irradiation, selected while considering the non-release of known disinfection by-products because of eventual reactions with the large organic and mineral load commonly present in the wastewater.

Bioaugmentation and phytoremediation wastewater treatment process as a viable alternative for pesticides removal: case of pentachlorophenol.

This study focused on the potential for pentachlorophenol removal by a biological process in secondary treated wastewater (STWW). The proposed process is a combined method of phytoremediation using a native plant, Polypogon maritimus and Lemna minor, and bioaugmentation using a fungus. The bioaugmentation process was performed by a fungal isolate capable of removing PCP, isolated from the compost. The identification of the fungus was performed by morphological, biochemical, and molecular methods. A biological treatment system by bioaugmentation and phytoremediation was set up to estimate the capacity of this process to eliminate a high concentration of PCP. physico-chemical parameters, such as pH, COD, and BOD were tested at experimentation times T0 (initial) and Tf (final). The concentration of PCP is controlled by the HPLC method. Thus, the growth of the fungus was determined by spectrophotometry and enumeration on the agar medium. The results obtained show that the isolated and selected fungus is identified by Penicillium Ilerdanum. The fungal strain used has a significant capacity for tolerance and elimination of PCP. The results of the physico-chemical parameters showed an improvement in the quality of wastewater after the treatment was carried out. The elimination of PCP came with a release of Common law- and an important decrease in the DOC value in the STWW. The results obtained show that the Polypogon treatment shows a significant elimination of PCP by a percentage of the order of 92.01% and 23.58 g. L- 1 chloride concentration. The macrophytes used showed a better ability to tolerate and eliminate PCP with an increase of chlorophyll and its longer sheets. Supplementary Information: The online version contains supplementary material available at 10.1007/s40201-023-00865-y.

Detection of Enteroviruses and SARS-CoV-2 in Tunisian Wastewater.

Monitoring the circulation of enteric viruses in environmental wastewater is a valuable tool for preventing the emergence of waterborne and food-borne diseases in humans. The detection of viruses was performed in five Tunisian wastewater treatment plants, three located in the Grand Tunis City (WWTP 1, WWTP 2, WWTP 3) and two in the Sahel of Tunisia (WWTP 4, WWTP 4), known as very developed and crowded zones, to assess the effectiveness of three biological wastewater treatment procedures namely natural oxidizing lagoons, rotating biodisks procedure, activated sludge procedure, and one tertiary sewage treatment using UV-C254 reactor for this enteric viruses' removal. Thus, 242 sewage samples were collected between June 2019 and May 2020 from different lines of wastewater treatment procedures implemented in the five wastewater treatment plants investigated. SARS-CoV-2 was analyzed using real-time multiplex reverse-transcription polymerase chain reaction (multiplex real-time RT-PCR) and enteroviruses using reverse-transcription polymerase chain reaction (RT-PCR). The enteroviruses detection showed 93% and 73% respective high frequencies only in the two WWTPs of the Grand Tunis (WWTP 1 and WWTP 2). SARS-CoV-2 was detected in 58% of the all wastewater samples collected from the five studied WWTPs with a respective dominance of N gene (47%), S gene (42%), RdRp gene (42%) and at last E gene (20%). These enteroviruses and SARS-CoV-2 detection were revealed in all steps of the wastewater treatment procedures, so poor virological quality is found at the exit of each biological and tertiary step of treatment investigated. For the first time in Tunisia, these results highlighted the enterovirus and SARS-CoV-2 detection with high rates, and the ineffectiveness of the biological and UV-C254 treatment implemented to remove these viruses. The preliminary results of SARS-CoV-2 circulation in Tunisian wastewater confirmed the wide positivity rate underlined by other works worldwide and allowed showing a move towards integrating wastewater as a way for this virus to spread in different areas and environments. So, this last result about SARS-CoV-2 circulation allowed us to caution about the strong probability of diffusion of this hazardous virus through water and sewage; despite its enveloped character and nature, as a labile and sensitive virus in these environments. Thus, establishing a national surveillance strategy is needed to improve the sanitary quality of treated wastewater and prevent public health problems related to these viruses in treated wastewater.

Effectiveness of combined tools: adsorption, bioaugmentation and phytoremediation for pesticides removal from wastewater.

The aim of this study was the comparison of two process in pentachlorophenol (PCP: 100 mg L-1) removal by combined process bioaugmentation-adsorption and bioaugmentation-phytoremediation in secondary treated wastewater (STWW). The phytoremediation procedure was conducted by using two plants such as Typha angustifolia and Schoenoplectus acutus, and the bioaugmentation procedure was operated by Pseudomonas putida HM 627618 as a plant growth promoting bacteria (PGPR). The adsorption process was performed by palm date activated carbon. The PCP monitoring was assessed by high performance liquid chromatography (HPLC) and the optical density determination at 600 nm (OD600). The performance of the two processes was observed by the determination of total bacteria, chlorophylls and physical and chemical analysis (COD, pH, conductivity, chloride, and organic carbon). The alfalfa seed germination test was conducted to assess the two operational performance procedures. According to the results obtained from the physical and chemical analysis of the treated STWW, there was no significant differences in the pH and in the EC content of the bioaugmentation-phytoremediation treatment, while a significant increase of the EC content was observed in the bioaugmentation-adsorption to 5.08 mS cm-1. The COD value significantly decreased up to 1320 mg L-1 in bioaugmentation-adsorption treatment (control value 2400 mg L-1) and 98 mg L-1 in bioaugmentation-phytoremediation treatment (control value 98 mg L-1). Microbial biomass monitoring of P. putida shows significant greater in both processes in the order of 9.18 and 7.01 Log CFU mL-1 for bioaugmentation-adsorption and bioaugmentation-phytoremediation, respectively. The chlorophyll content in Typha angustifolia and Schoenoplectus acutus significantly decreased after 144 h with the exception of the chlorophyll a content of Schoenoplectus acutus in which the content increased up to 3.31 mg mL-1. Comparing the performance of these two treatments, it was found according to HPLC analysis that the bioaugmentation-adsorption process was more efficient in removing about 97% of PCP after 48 h, against around 90% of PCP after 72 h for the phytoremediation-bioaugmentation. The alfalfa seeds showed a germination rate after the 5th day of incubation of 100% and 95%, respectively for the PCP-non-contaminated and treated STWW, while for wastewater containing PCP the germination was totally inhibited.

This paper describing sensitive methods of combined bioaugmentation-phytoremediation and bioaugmentation-adsorption for pentachlorophenol (PCP) depletion in wastewater. The novelty is the choice of a macrophyte Typha angustifolia and Schoenoplectus acutus in constructed wetland fixed in clay matrix. The two-selected plants are still used for the elimination of heavy metals but not for pesticide in wastewater. Also, the combined process bioaugmentation-adsorption was not tested in other researches. On the other side, in this study, the phytoremediation technique combined with bacteria positively affected the plants activity in order to promote pollutant remediation. Hence, the Pseudomonas putida HM 627618 in wastewater with the macrophyte presence or date stone adsorbent have a great capacity to reduce this pollutant (PCP) by improving the bioremediation process.

Widespread of the Vienna/Hungarian/Brazilian CC8-ST239-SCCmec III MRSA clone in patients hospitalized in the Tunisian Burn and Traumatology Center.

The emergence and spread of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals is a major global public health concern. The current study sought to characterize 25 MRSA clinical isolates collected in a Tunisian hospital from December 2015 to September 2016, with the genetic lineages, virulence factors, and antibiotic resistance mechanisms determined for these isolates. Three spa-types were detected: t037 (23 isolates), t932, and t2235 (one isolate each). Isolates were ascribed to agr I (n = 20), agr II (n = 1), with four nontypeable isolates. Depending on sequence type (ST), the 25 MRSA isolates were assigned to two clonal complexes (CC8 and CC5), with a predominance of the lineage ST239-CC8 (n = 24; 96%). All isolates belonging to CC8 had the SCCmec type III, while the unique CC5 isolate had SCCmec type IV. Antimicrobial susceptibility testing revealed high levels of resistance to aminoglycosides, tetracycline, ciprofloxacin and rifampicin for the majority of isolates belonging to the ST239-CC8 lineage. The ST149-CC5 isolate was susceptible to non-ß-lactam antibiotics. One isolate harbored the tsst-1 gene (4%); however, lukS/LukF-PV, eta and etb genes were not detected. The MDR ST239-CC8 clone would seem to be widespread in this hospital. Therefore, a rigorous hygienic control system is urgently required.

Co-occurrence of antibiotic and metal resistance in long-term sewage sludge-amended soils: influence of application rates and pedo-climatic conditions.

Urban sewage sludge (USS) is increasingly being used as an alternative organic amendment in agriculture. Because USS originates mostly from human excreta, partially metabolized pharmaceuticals have also been considered in risk assessment studies after reuse. In this regard, we investigated the cumulative effect of five annual USS applications on the spread of antibiotic-resistant bacteria (ARB) and their subsequent resistance to toxic metals in two unvegetated soils. Eventually, USS contained bacterial strains resistant to all addressed antibiotics with indices of resistance varying between 0.25 for gentamicin to 38% for ampicillin and azithromycin. Sludge-amended soils showed also the emergence of resistome for all tested antibiotics compared to non-treated controls. In this regard, the increase of sludge dose generally correlated with ARB counts, while soil texture had no influence. On the other hand, the multi-antibiotic resistance (MAR) of 52 isolates selected from USS and different soil treatments was investigated for 10 most prescribed antibiotics. Nine isolates showed significant MAR index (≥ 0.3) and co-resistance to Cd, As and Be as well. However, events including an extreme flash flood and the termination of USS applications significantly disrupted ARB communities in all soil treatments. In any case, this study highlighted the risks of ARB spread in sludge-amended soils and a greater concern with the recent exacerbation of antibiotic overuse following COVID-19 outbreak.

Effects of heavy metals on growth and biofilm-producing abilities of Salmonella enterica isolated from Tunisia.

This study aims to test the toxicity of some metallic elements on Salmonella enterica strains and their power to grow and to develop a biofilm to overcome this environmental stress. From 50 selected strains of Salmonella, 70% belong to the Kentucky serotypes that is the most frequent one, followed by the other serotypes such as Amsterdam 6%, anatum 4%, derby 4% Enteritidis 4%, Zanzibar 4%, typhyrimium 2%, gallinaruim 2%, inbondaka 2% and Newport 2%. All the strains have presented the invA invasion gene involved in the virulence and Salmonella infection. Genotypic BOX-PCR analysis of these strains showed 18 profiles, with a discrimination index of 0.93. The Salmonella growth has mainly revealed that the variation of the rates of different metallic elements showed a significant influence on the Salmonella growth. The qualitative, quantitative study and biofilm tubes showed that 40% of the strains have a strong capacity to form biofilm, and the wild-type phenotypes (RDAR; rigid film; Strong), This phenotype varies according to the nature and the concentration of the metal (0.1 mM-1 mM) considered. In the presence of copper, zinc, cobalt, and chromium, the Salmonella strains showed a potent capacity to form a biofilm with a slight variation in the wild-type phenotype. However, when chromium rates increased, Salmonella loses the RDAR morphotype. Addition of mercury and cadmium in the growth medium reduced the production of Salmonella biofilm by around 14 and 15%, respectively, if compared with the control free of metals.

High-Resolution Genotyping Unveils Identical Ampicillin-Resistant Enterococcus faecium Strains in Different Sources and Countries: A One Health Approach.

Multidrug-resistant (MDR) Enterococcus faecium (Efm) infections continue to increase worldwide, although epidemiological studies remain scarce in lower middle-income countries. We aimed to explore which strains circulate in E. faecium causing human infections in Tunisian healthcare institutions in order to compare them with strains from non-human sources of the same country and finally to position them within the global E. faecium epidemiology by genomic analysis. Antibiotic susceptibility testing was performed and transfer of vancomycin-vanA and ampicillin-pbp5 resistance was performed by conjugation. WGS-Illumina was performed on Tunisian strains, and these genomes were compared with Efm genomes from other regions present in the GenBank/NCBI database (n = 10,701 Efm genomes available May 2021). A comparison of phenotypes with those predicted by the recent ResFinder 4.1-CGE webtool unveiled a concordance of 88%, with discordant cases being discussed. cgMLST revealed three clusters [ST18/CT222 (n = 13), ST17/CT948 strains (n = 6), and ST203/CT184 (n = 3)], including isolates from clinical, healthy-human, retail meat, and/or environmental sources in different countries over large time spans (10-12 years). Isolates within each cluster showed similar antibiotic resistance, bacteriocin, and virulence genetic patterns. pbp5-AmpR was transferred by VanA-AmpR-ST80 (clinical) and AmpR-ST17-Efm (bovine meat). Identical chromosomal pbp5-platforms carrying metabolic/virulence genes were identified between ST17/ST18 strains of clinical, farm animal, and retail meat sources. The overall results emphasize the role of high-resolution genotyping as provided by WGS in depicting the dispersal of MDR-Efm strains carrying relevant adaptive traits across different hosts/regions and the need of a One Health task force to curtail their spread.

Macrophyte and indigenous bacterial co-remediation process for pentachlorophenol removal from wastewater.

This study has contributed in the description of bioaugmentation-phytoremediation efficiency process using Typha angustifolia concerning PCP tolerance and removal from wastewater. Samples of wastewater were collected from industrial wastewater plants, namely row wastewater effluent "E.WW", primary wastewater "P.WW", secondary wastewater "S.WW", clarified wastewater "AC.WW". These effluents were spiked with PCP at different rate (100, 500, and 1000 mg.L-1), physical, chemical and biological properties were monitored. A second experiment was set up in order to check the efficiency of phytoremediation treatments of the different effluents artificially contaminated with 200 mg.L-1 PCP after 20 days lab scale experiment. An important PCP removal by indigenous bacteria was showed in S. WW with values from 1000 to 72.2 mg.L-1 from T0 (start of the experiment) to TF (end of the experiment), respectively. Phytoremediation process allowed a decrease of PCP rate from 200 to 6.4 mg.L-1, a decrease of chloride content from 14.0 to 4.0 mg.L-1 in S.WW samples was observed. Furthermore, a significant increase of bacterial number in S.WW and AC.WW to 1.700 × 106 and 1.450 × 106 CFU.mL-1, respectively was observed. In addition, the DGGE analysis showed that after bioaugmentation-phytoremediation treatments, the highest species richness and relative abundance in wastewater effluent was observed. Novelty statement Pentachlorophenol (PCP) is one of highly toxic of polychlorophenols and required to continuously monitor in environment. This paper presenting a sensitive method phytoremediation and bioaugmentation for PCP biotransformation in wastewater. The novelty is the choice of a macrophyte Typha angustifolia, which is still used for the elimination of heavy metals but it not used for pesticide and pollutant removal in wastewater. Also, there are different analysis that was performed in order to check phyto-technique process (DGGE and HPLC). On the other side, in this study, the phyto-techniques with Typha angustifolia positively affected intrinsic microorganisms in order to promote pollutant remediation. So, the intrinsic microorganisms in wastewater with the macrophyte presence have a great capacity to reduce this pollutant and improve the bioremediation process.

Combined bioaugmentation and biostimulation techniques in bioremediation of pentachlorophenol contaminated forest soil.

Pentachlorophenol (PCP) is quite persistent in the environment and severely affects different ecosystems including forest soil. The main objective of this work was to study different bioremediation processes of artificially PCP (100 mg kg-1) contaminated forest soil (Sc). In fact, we used bioaugmentation by adding two different bacterial consortia B1 and B2, biostimulation procedures by amendments based on forest compost (FC), municipal solid waste compost (MC), sewage sludge (SS), and phosphate, and their combined treatments. Soil physical and chemical properties, residual PCP, soil microbial biomass carbon, soil respiration and some enzymatic activities at zero time and after 30 d of incubation, were evaluated. A net reduction of PCP, 71% of the initial concentration, after 30 d-incubation occurred in the sample Sc+B1+FC, as the best performance among all treatments, due to natural attenuation, immobilization of PCP molecules in the forest soil through organic amendments, and the action of the exogenous microbial consortium B1. The single application of FC or B1 led to a depletion of PCP concentration of 52% and 41%, respectively. Soil microbial biomass carbon decreased in PCP contaminated soil but it increased when organic amendment also in combination with microbial consortia was carried out as bioremediation action. Soil respiration underwent no changes in contaminated soil and increased under FC based bioremediation treatment. These results demonstrate that the combined treatments of biostimulation and bioaugmentation might be a promising process for remediation of PCP contaminated soil.

High rates of antibiotic resistance and biofilm production in Escherichia coli isolates from food products of animal and vegetable origins in Tunisia: a real threat to human health.

The aim of this study was to compare the antibiotic susceptibility of eighty Escherichia coli isolates from vegetables and food products of animal origin in Tunisia, and to study their genes encoding antibiotic resistance and in vitro biofilm forming capacity. Antimicrobial susceptibilities were determined, as well as PCR investigation of genes associated with antibiotic resistance. Biofilm formation was tested using four different methods: the microtiter plate-, MTT-staining-, XTT-staining-, and the Congo Red Agar assays. High antibiotic resistance rates were observed for amoxicillin (68.7%), amoxicillin/clavulanic acid (73.7%), gentamicin (68.7%), kanamycin (66.2%), nalidixic acid (36.2%), streptomycin (68.7%) and tetracycline (35%). The majority of isolates was multidrug resistant and biofilm producer. MTT testing showed that vegetables isolates were significantly higher biofilm producers compared to foods of animal origins. This study showed that E. coli isolates from food products were reservoirs of genes encoding antibiotic-resistance and have a high propensity to produce biofilm.

Study of the diversity of 16S-23S rDNA internal transcribed spacer (ITS) typing of Escherichia coli strains isolated from various biotopes in Tunisia.

We investigated the 16S-23S rRNA intergenic spacer region (ISR)-PCR and the phylogenetic PCR analyses of 150 Escherichia coli isolates as tools to explore their diversity, according to their sampling origins, and their relative dominance in these sampling sources. These genetic markers are used to explore phylogenetic and genetic relationships of these 150 E. coli isolates recovered from different environmental sources (water, food, animal, human and vegetables). These isolates are tested for their biochemical pattern and later genotyped through the 16S-23S rRNA intergenic spacer PCR amplification and their polymorphism investigation of PCR-amplified 16S-23S rDNA ITS. The main results of the pattern band profile revealed one to four DNA fragments. Distributing 150 E. coli isolates according to their ITS and using RS-PCR, revealed four genotypes and four subtypes. The DNA fragment size ranged from 450 to 550 bp. DNA band patterns analysis revealed considerable genetic diversity in interspecies. Thus, the 450 and 550 bp sizes of the common bands in all E. coli isolates are highly diversified. Genotype I appeared as the most frequent with 77.3% (116 isolates), genotype II with 12% (18 isolates); genotype III with 9.7% (14 isolates), and the IV rarely occurred with 4% (2 isolates). Distributing the E. coli phylogroups showed 84 isolates (56%) of group A, 35 isolates (23.3%) of group B1, 28 isolates (18.7%) of group B2 and only three isolates (2%) of group D.

Removal of pentachlorophenol from contaminated wastewater using phytoremediation and bioaugmentation processes.

The phytoremediation procedure was conducted by Lemna gibba (L) and Typha angustifolia (T) and the bioaugmentation procedure used P. putida HM627618. The ability of the selected P. putida HM627618 to tolerate and remove PCP (200 mg L-1) was measured by high performance liquid chromatography analysis and optical density at 600 nm. Five different experiments were conducted in secondary treated wastewater for PCP testing removal (100 mg L-1) including two phytoremediation assays (T + PCP; L + PCP), three bioaugmentation-phytoremediation assays (T + B + PCP; L + B + PCP; L + T + B + PCP) and a negative control assay with PCP. Various analytical parameters were determined in this study such as bacterial count, chlorophylls a and b, COD, pH and PCP content. The main results showed that the average PCP removal by P. putida HM627618 was around 87.5% after 7 days of incubation, and 88% of PCP removal was achieved by treatment (T + B) after 9 days. During these experiments, pH, COD and chloride content showed a net increase in all treatments. The chlorophylls a and b in case of (T) and (L) Chlorophylls a and b for T and L phytoremediation showed a decrease with a value less than 10 µg/mg of fresh material after 20 days of cultivation.

Bacterial consortium biotransformation of pentachlorophenol contaminated wastewater.

The aims of this study were (i) to compare PCP removal (100 mg L-1) by two bacterial consortia B1 and B2 in sterile wastewater (STWW) and liquid mineral medium (MSM), (ii) PCP effect in biofilm formation and antimicrobial susceptibility. PCP removal was measured by high-performance liquid chromatography (HPLC) during 168 h at 30 °C. Biofilm formation was assessed with two approaches: Congo Red Agar and Microtiter-plate. Antimicrobial susceptibility was determined by the agar disc diffusion technique. The results showed that the PCP removal for consortium B1 and B2 after 168 h was 70 and 97.5% in STWW; 62.2 and 85.5% in MSM, respectively. In addition, PCP addition showed an increase in biofilm development especially for B2 consortium around 3.5 nm in 100 mg L-1 PCP. PCP added in the Muller Hinton (MH) medium and Gentamicin disc showed a clear increase in diameter of cell lysis around 2 to 4.5 cm.

Surfactant efficiency on pentachlorophenol-contaminated wastewater enhanced by Pseudomonas putida AJ 785569.

This study aims to evaluate the effect of three surfactants on the removal of PCP (800 mg L-1) from Secondary Treated Wastewater (STWW) by Pseudomonas putida AJ 785569. The effect of surfactants [sodium lauryl sulfate (SDS) as anionic, Tween 80 (TW80) as non-anionic and cetyltrimethylammonium bromide (CTAB) as cationic] is tested about the three following aspects: (1) bacterial growth, (2) bacterial biofilm formation or development and (3) PCP rate removal. The results showed that strain P. putida AJ 785569 could adsorb around 30 mg L-1 and remove 600 mg L-1 of PCP within 168 h of incubation. The SDS developed the growth of bacteria and the removal of PCP. This PCP removal in mineral salt medium (MSM) is around 760 mg L-1 (95% degradation) higher than the ones registered with CTAB and TW80 with a value 506.75 (63% degradation) and 364.1 mg L-1 (45% degradation), respectively. The obtained results of chloride concentration showed an important relation with PCP removal during incubation with an important value. Monitoring the development of bacterial biofilm, in MSM medium added with PCP (100 mg L-1) by strain P. putida AJ 785569, showed a significant increase in the optical density value from 0.9 to 4 at λ = 595 nm, a modification of strain P. putida AJ 785569's morphotype, density and color colonies.

Exhaustion of pentachlorophenol in soil microcosms with three Pseudomonas species as detoxification agents.

Pentachlorophenol (PCP) is a toxic compound, which is widely used as a wood preservative product and general biocide. It is persistent in the environment and has been classified as a persistent organic pollutant to be reclaimed in many countries. Bioremediation is an emerging approach to rehabilitating areas polluted by recalcitrant xenobiotics. In the present study, we evaluated the potential of three strains of Pseudomonas (P. putida S121, P. rhizophila S211, and P. fuscovagiceae S115) as bioremediation agents in depletion and detoxification of PCP in soil microcosms. PCP removal was effectively optimized using a central-composite experimental design and response surface methodology (RSM). The optimum conditions for maximum PCP removal yield (85 ± 5%) were: 500 mg/kg PCP concentration, 108 UFC/g soil inoculum size of each strain and 55 days incubation period. The bacterial strains, P. putida, P. rhizophila, and P. fuscovagiceae, showed good capability to tolerate and degrade PCP so that they could be successfully used in synergistic effect to treat PCP polluted soils.

Inactivation of Hepatovirus A in wastewater by 254 nm ultraviolet-C irradiation.

Hepatovirus A is known as a waterborne and foodborne virus that can be transmitted from one person to another through contaminated water and raw food. Therefore, it is necessary to survey the circulation of this type of enteric virus in the wastewater to prevent prospective outbreaks. Wastewater samples collected from WWTP El Menzeh I and Charguia I have been the subject for physicochemical, bacteriological (MPN) and virological analyses. Hepatovirus A (HAV) detection was carried out using the standard reverse transcription-polymerase chain reaction (RT-PCR). Hepatovirus A was detected respectively in 62% (63/102) and 66% (92/140) of the collected wastewater samples at El Menzeh I and Charguia I WWTPs. The treated effluent by natural oxidizing lagoon procedure was characterized by a poor physical-chemical and virological qualities but with excellent bacteriological quality. Consequently, this effluent is not suitable to be recycled and reused in agriculture or even dismissed in the environment. The treated sewage by activated sludge and rotating biodisk procedures turned out to be of a very good physical-chemical quality but with a poor bacteriological and virological quality. After tertiary UV-C254 nm irradiation, the faecal indicator bacteria concentration was mostly reduced and removed. These findings confirmed the need for improvement and upgrade of the treatment processes used in these two studied sewage purification plants and the necessity of implementation and establishment of a proper national virological standard to control the circulation rates of enteric viruses in Tunisian municipal wastewater.

Genetic characterization of extended-spectrum ß-lactamase-producing Enterobacteriaceae from a biological industrial wastewater treatment plant in Tunisia with detection of the colistin-resistance mcr-1 gene.

This study evaluated the occurrence of extended-spectrum ß-lactamases (ESBL) and associated resistance genes, integrons, and plasmid types, as well as the genetic relatedness of enterobacterial isolates in the wastewater treatment plant (WWTP) of La Charguia, Tunis City (Tunisia). A total of 100 water samples were collected at different points of the sewage treatment process during 2017-2019. Antimicrobial susceptibility was conducted by the disc-diffusion method. blaCTX-M, blaTEM and blaSHV genes as well as those encoding non-ß-lactam resistance, the plasmid types, occurrence of class1 integrons and phylogenetic groups of Escherichia coli isolates were determined by PCR/sequencing. Genomic relatedness was determined by multi-locus sequence typing (MLST) for selected isolates. In total, 57 ESBL-producer isolates were recovered (47 E. coli, eight Klebsiella pneumoniae, 1 of the Citrobacter freundii complex and 1 of the Enterobacter cloacae complex). The CTX-M-15 enzyme was the most frequently detected ESBL, followed by CTX-M-27, CTX-M-55 and SHV-12. One E. coli isolate harboured the mcr-1 gene. The following phylogroups/sequence types (STs) were identified among ESBL-producing E. coli isolates: B2/ST131 (subclade-C1), A/ST3221, A/ST8900, D/ST69, D/ST2142, D/ST38, B1/ST2460 and B1/ST6448. High numbers of isolates harboured the class 1 integrons with various gene cassette arrays as well as IncP-1 and IncFIB plasmids. Our findings confirm the importance of WWTPs as hotspot collectors of ESBL-producing Enterobacteriaceae with a high likelihood of spread to human and natural environments.