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Aquatic environments are crucial hotspots for the dissemination of antibiotic resistant microorganisms and resistance genes. Thus, the purpose of this study was to investigate the occurrence and the genetic characterization of cefotaxime-resistant (CTXR) Enterobacteriaceae at a Tunisian semi-industrial pilot plant with biological treatment (WWPP) and its receiving river (Rouriche River, downstream from WWPP) located in Tunis City, during 2017–2018. We collected 105 and 15 water samples from the WWPP and the Rouriche River, respectively. Samples were screened to recover ESBL-producing Enterobacteriaceae (ESBL-E) and isolates were characterized for phenotype/genotype of antimicrobial resistance, integrons, plasmid types and molecular typing (multilocus sequence typing, MLST). Among 120 water samples, 33 and 4 contained ESBL-producing E. coli and K. pneumoniae isolates, respectively. Most isolates were multidrug resistant and produced CTX-M-15 (28 isolates), CTX-M-1 (4 isolates), CTX-M-55 (2 isolates), CTX-M-27 (one isolate), SHV-12 (one isolate) and VEB beta-lactamases (one isolate). All K. pneumoniae were CTX-M-15-positive. Four colistin-resistant isolates were found (MIC 4–8 μg/ml), but they were negative for the mcr genes tested. Class 1 integrons were detected in 21/25 trimethoprim/sulfamethoxazole-resistant isolates, and nine of them carried the gene cassette arrays: aadA2 + dfrA12 (n = 4), aadA1 + dfrA15 (n = 2), aadA5 + dfrA17 (n = 2) and aadA1/2 (n = 1). The IncP and IncFIB plasmids were found in 30 and 16 isolates, respectively. Genetic lineages detected were as follows: E. coli (ST48-ST10 Cplx, ST2499, ST906, ST2973 and ST2142); K. pneumoniae: (ST1540 and ST661). Our findings show a high rate of CTX-M-15 and high genetic diversity of ESBL-E isolates from WWPP and receiving river water.

Several mathematical models are developed to explain microorganism responses to ultraviolet irradiation. During this study, Pseudomonas aeruginosa was taken as a microorganism model. The experimental results obtained during a batch UV-reactor established that the disinfection kinetics is far from being uniform over the batch. Application of the first-order Chick-Watson model, in its original form or modified form, to validate the speed changes over the disinfection process, does not report any significantly improved results. Application of the Collins-Selleck model fits better the kinetic curves of UV disinfection of wastewater for all the tested strains of P. aeruginosa. At the same time, various continuous alternative reactors are checked, by accounting for the fluid flow hydrodynamics, that is, a perfectly mixing reactor without and with short circuits in the flow stream, or without and with dead zone inside the reactor, a plug flow reactor, and a series of perfectly mixed reactors. The results indicate that the succession of 4 perfectly mixed reactors is the best alternative to the process scale-up. Key words: Disinfection, hydrodynamics, kinetics model, reactor of disinfection, treated wastewater, UV radiation.

The aim of this paper was to propose a modeling system of water ultraviolet (UV) disinfection. Results reveal that application of the model of Chick-Watson in its original form or modified are not representative of the kinetics of disinfection. For this reason, the application of a new kinetic model of Collins-Selleck in UV inactivation of Pseudomonas aeruginosa in secondary wastewater appeared to be the best applied model. The modeling of the reactivation process at range of 7.5-50°C temperature was shown. First-order saturation does not fit the obtained data in photoreactivation; a modification of the model is proposed coinciding with the classical logistic equation. To better explain the process of inactivation, we have assumed that the action of disinfectant on the survival of lonely microorganisms is faster than its action on suspended solids protected or agglomerated to each other. For this reason, the application of a new kinetic model by introducing a third factor reflecting the influence of suspended solids in water on disinfection kinetics appeared to be determinant for modeling UV inactivation of P. aeruginosa in secondary treated wastewater. Key words: Secondary wastewater, ultraviolet (UV) disinfection, modeling, photoreactivation. Pseudomonas aeruginosa, suspended solids.

The present study deals with the isolation, the identification, and the characterization of heavy metal-resistant bacteria selected from several polluted sources with the aim of using them as bioremediation agents of contaminated sites by heavy metals. Four protocols were used in a different way to select the maximum of isolates resistant to heavy metals. Isolates obtained, according to the fourth selection protocol by using the free nutrients medium, showed the highest values of minimum inhibitory concentration (MIC), and it can be deduced from this last result that when the bacteria are subjected to stressful conditions they develop a system that will enable them to withstand high levels of heavy metals. The study of the MIC at all resistant isolates allowed determining that the order of toxicity of the different metals was as follows: Cu > Cr > Cd > Ni > Zn > Co. These metals might exert varying effects depending on their adsorption and their complexation capacities to the bacterial cell wall. ...

Sewage disinfection has the primary objective of inactivating pathogenic organisms to prevent the dissemination of waterborne diseases. This study analyzed individual disinfection, with chlorine alone, ultraviolet radiation alone, and a combined disinfection process (chlorine-UV radiation). Pseudomonas aeruginosa ATCC 15442, Escherichia coli ATCC 11229, Salmonella typhi ATCC 14028, and Clostridium perfringens were selected to evaluate the efficiency of different disinfection processes. The aim of the present study was to characterize the kinetics of chlorine (as NaHOCl) consumption, to evaluate responses of these bacterial species to the chlorination, the ultraviolet (UV) radiation, and the chlorine/UV disinfection processes in secondary wastewater using a batch laboratory reactor. Another target of this work was to study the modeling of the kinetic of water disinfection by chlorination and/or UV irradiation. Two kinetic models (Chick-Watson and Hom) were tested as to ability to scale disinfection of these bacterial species by different ultraviolet and/or chlorine doses. The results of the kinetics of chlorine consumption showed that monochloramines and trichloramines were the most important forms of residual chlorine as compared to free chlorine and dichloramines. The kinetics of inactivation of all examined bacterial strains showed that the application of the model of Hom in its original form was not representative of this kinetics of inactivation. Modification of this model, considering an initial decline of bacteria during the contact of water with chlorine, improved the results of the model. By the same, results revealed that the involved processes of UV irradiation were too complex to be approached by a simplified formulation, even in the case of specific strains of microorganisms and the use of nearly constant UV radiation intensity. In fact, the results have pointed out that the application of the Chick-Watson law is known to be inadequate to describe microbial inactivation with tailing or shoulder behavior. However, the UV kinetic studies also revealed that the first instants of exposure (2 to 10 s) to a UV intensity of 5 to 8 mW.cm-2 appeared as the deciding factors in disinfection with UV irradiation. It was shown that the combination of chlorine with UV yielded additive effects on the inactivation of bacterial strains and spectacular disinfection efficiency at a short contact time and less dose. In contrast to most studies, UV irradiation alone and chlorine alone exhibited low disinfection efficiency in inactivation of tested bacterial strains.

The purpose of this research stream is to evaluate the kinetics of bacterial strains of Pseudomonas aeruginosa, to establish the influence of UV doses on the kinetics of disinfection, to study UV-resistant strains of Pseudomonas aeruginosa, and to underline the influence of suspended solids on the inactivation kinetics of these strains. Furthermore, and due to the lack of readily available information about the influence of temperature on microorganism inactivation processes subsequent to inactivation with UV radiation, a series of batch studies were performed at 5℃, 25?C, 37.5℃ and 50℃. This paper investigates the impact of UV irradiation on bacterial strains of P. aeruginosa inactivation in both primary and secondary wastewater effluents and to show the influence of filtration in the process of disinfection of water by UV irradiaton. Our results indicate that the effect of temperature within the normal operating range of most treatment plants, i.e., 25℃ to 37.5℃, was found to be not statistically significant on the kinetics of the UV disinfection process. However, the kinetics of the UV disinfection process was highly affected by system operating at extreme temperatures, i.e., at 10 and 50℃. In a temperature range of 25℃ to 37.5℃, the inactivation of P. aeruginosa strains varied according to the incubation time and did not exceed 4 U-Log. Consequently, having more than 102 organisms/100ml of P. aeruginosa in treated wastewater would cause serious health and environmental problems. Low inactivation was observed when the operating temperature was reduced to 10℃ regardless of the incubation time tested. In contrast, a considerable increase in the inactivation rate was noted when the temperature of the disinfected wastewater was increased to 50℃. Hence, as 5℃ and 50℃ are outside the operating range of most treatment plants, as in our country, Tunisia, it is reasonable to assume that the effect of temperature change on the kinetics of UV disinfection process is negligible.

The main objective of this investigation was to study the enzymatic activities of Actinomycetales strains isolated from an agricultural soil amended with different amounts of municipal solid waste compost (MSWC) or farmyard manure (FM). For this purpose, the hydrolytic activities of carboxymethyl cellulase, xylanase, pectinase, amylase, chitinase and protease were tested for 75 isolates of Sterptomyces, Amycolatopsis and Nocardioides from different sources (unamended soil, amended soil with FM or MSWC, FM and MSWC) at temperature ranging between 30 and 50°C. It was shown that the highest rate of enzymes producer’s strains was registered at 30°C, and decreased gradually to annul at 50°C, with the exception of the MSWC strains origin. It was also shown that the percentage of strains producers of enzymes isolated from soil amended with MSWC appeared higher than the one registered for those isolated from control and amended with FM soils. Application of MSWC increases the number of enzymes producer-actinomycetes in the soil and then it improves its fertility.

The main objective of this investigation was to study the Actinomycetales community in a Tunisian agricultural soil amended with different amounts of municipal solid waste compost (MSWC) or farmyard manure (FM). It was shown, by using the 16S rDNA polymerase chain reaction—restriction fragment length polymorphism and sequencing methods for Actinomycetales isolates, tow major clusters. The cluster SI is composed of tow related families: Streptomycetaceae, dominant family and Pseudonocardiaceae. However, SII is composed of Nocardioidaceae family. The application of FM or MSWC increases the Actinomycetales number and affects their diversity by inhibition, stimulation or contribution of some species in soil. It was also shown, by the application of molecular techniques to soil samples using selective primers, that these three groups of Actinomycetales remain dominant even after 5 years of organic amendment application in soil.

In order to test the ability to produce antibacterial substances within marine bacteria, prior to select potential probiotics for use in shellfish farming, we targeted a large collection of bacterial isolates (132 strains), brought from the clamRuditapes decussatus and 37 reference strains. First, we proceeded to their biochemical identification and the screening of antibiotic resistance profiles. Else, we investigated their inhibitory activityin vitro against several fish and shellfish pathogens, using two methods: the double-layer agar and the direct simultaneous antagonism methods. The results showed high frequencies of inhibitory producing bacteria (IPB) within the isolates. These bacteria (25%) were aerobic mesophylic bacteria belonging to various bacterial groups: 33.7% oxidase-positive Gram-negative bacteria, 7.4%Enterobacteriaceae and 28% lactic acid bacteria. Besides this group, nine strains produced strong inhibition effect. These bacteria belonged to:Aeromonas hydrophila, Aeromonas sobria, Pseudomonas cepacia, Vibrio sp,Serratia liquefaciens andLactobacillus rhamnosus. They were active against pathogenic bacteria belonging to the genera:Aeromonas, Pseudomonas andVibrio. These potential probiotics were submitted to further investigations prior to their introduction in larval shellfish farming.

The ubiquitous coexistence of heavy metals and organic contaminants was increased in the polluted soil and phytoremediation as a remedial technology and management option is recommended to solve the problems of co-contamination. Growth of Zea mays L and pollutant removal ability may be influenced by interactions among mixed pollutants. Pot-culture experiments were conduced to investigate the single and interactive effect of cadmium (Cd) and pentachlorophenol (PCP) on growth of Zea mays L, PCP, and Cd removal from soil. Growth response of Zea mays L is considerably influenced by interaction of Cd and PCP, significantly declining with either Cd or PCP additions. The dissipation of PCP in soils was notably affected by interactions of Cd, PCP, and plant presence or absence. At the Pentachlorophenol in both planted and non-planted soil was greatly decreased at the end of the 10-week culture, accounting for 16-20% of initial extractable concentrations in non-planted soil and 9-14% in planted soil. With the increment of Cd level, residual pentachlorophenol in the planted soil tended to increase. The pentachlorophenol residual in the presence of high concentration of Cd was even higher in the planted soil than that in the non-planted soil.

This paper aimed to the study of microbiological quality of wastewater effluents issued from a sand filter system used as a secondary treatment; this sand filter was coupled to UV irradiation system as a tertiary treatment to assume their possible application as alternatives to chemical disinfection of municipal wastewater before agronomic utilization. For that reason, an infiltration basin of 100 m² of area equipped with two wells of sampling, in the center and in the periphery, has been constructed on the irrigated perimeter of Dissa in Gabes (south east of Tunisia). A comparative study was undertaken concerning the quality of effluents obtained from the sand filter as secondary effluent and from the UV irradiation disinfection system as a tertiary municipal wastewater treatment. Both the combined systems gave effluent of excellent microbiological quality (almost total absence of E. coli, faecal coliforms and P. aeruginosa). However, if the microbiological quality of effluent remained constant in the case of a sand filter system which depends only on the quality of the influent, while with the UV-disinfection process, microbiological quality of tertiary effluent was shown to depend on the quality of secondary effluent. The monitoring of the UV irradiation device using a UV dose of around 96 mW.s.cm-2 and corresponding to an exposure time of 16 sec, for the secondary effluent at the outlet of Sand Filter, showed that the average bacterial inactivation rate was around 3 U-Log for E. coli, faecal coliforms and P. aeruginosa, respectively. Therefore, the average concentration remaining in the effluent at the output of the UV reactor, was less than a 100 cfu/ 100 ml for E. coli and faecal coliforms, and less than a 100 bacteria/ 100 ml for P. aeruginosa. These values coincided with the range recommended by several standardized international guidelines.

This paper deals with the study of bacteriological quality of effluents that have undergone consecutively different macrofiltration system (pressure sand filter or disc filter used as a secondary treatment) and UV254 irradiation process (used as a tertiary treatment). These two successive systems of treatment were evaluated to determine their possible application as commonly alternatives to the conventional system of wastewater treatment and disinfection before wastewater reuse. They both combined systems of wastewater treatment released effluent of excellent bacteriological quality, with almost total absence of feacal coliforms, of E. coli and of P. aeruginosa). However, if the bacteriological quality of the effluent remained constant in the case of macrofiltration system (disc filter or pressure sand filter); the UV disinfection process showed to deeply depend on the quality of effluent, particularly with regard to UV transmittance. The daily bacteriological monitoring of the secondary effluent at the exit of the pressure sand filter by UV reactor and by using a dose of 96 mJ/cm(2), corresponding to an exposure of 16 min, showed an average rate of inactivation of around 3 U-Log, for feacal coliforms, E. coli and P. aeruginosa, respectively. Therefore, the average bacterial concentration remaining in the water at the exit of the UV reactor is less than 1000 cfu/100 ml for feacal coliform and E. coli. For P. aeruginosa, the remaining number is less than 100 bacteria/100 ml. These two last values coincide substantially with the range recommended by several standardized international guidelines. Therefore, numerous authors reported that P. aeruginosa is very resistant to UV irradiation compared to the other bacterial indicators. In contrast, our study revealed that feacal coliforms and E. coli were more UV light resistant than P. aeruginosa. This finding could be explained by the fact that E. coli and feacal coliform forms aggregates in the treated effluent, while P. aeruginosa exists either as discrete cells or as cell pairs.

Actinomycetes with the ability to degrade natural polysaccharides were isolated during a screening programme from soil, farmyard manure and municipal solid waste compost. One of the most potent isolates was identified as Streptomyces sp. MSWC1 using morphological and biochemical properties along with 16S rDNA partial sequence analysis. The highest enzyme production by Streptomyces was observed for the xylanase and chitinase activity on different carbon sources with an optimum of 12,100 IU ml—1 and 110 IU ml—1 at 3 days’ culture on 1% of xylan and chitin, respectively. To meet the demand of industry, low-cost medium is required for the production of hydrolases by Streptomyces sp. Strain MSWC1 grown on manure, compost, and a natural carbon source was used to evaluate the re-utilisation of biological wastes for the production of value-added products. Despite the presence of a high amount of toxic heavy metals in the compost, Streptomyces produced interesting enzymes that have been biochemically characterized.

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.