Potential of Trametes trogii culture fluids and its purified laccase for the decolorization of different types of recalcitrant dyes without the addition of redox mediators

Trametes trogii BAFC 463 culture fluids (containing 110 U ml⁻¹ laccase; 0.94 U ml⁻¹ manganese peroxidase), as well as its purified laccase were capable of decolorizing azoic, indigoid, triphenylmethane, anthraquinonic and heterocyclic dyes, in the absence of redox mediators. Six dyes: RBBR, Indigo Carmine, Xylidine, Malachite Green, Gentian Violet and Bromophenol Blue were almost completely degraded (more than 85% decolorization after 1 d) by either laccase or T. trogii itself in culture, proving the role of the enzyme in dye decolorization. The purified laccase also decolorized 65% of Fast Blue RR and 30% of Azure B and Methylene Blue after 24 h. The use of redox mediators significantly increased the decolorization rates (90% decolorization of Azure B after 1 h). 1-hydroxybenzotriazole resulted the best redox mediator, but the natural mediator p-hydroxybenzoic acid also demonstrated its efficiency for dye decolorization. Due to their ability to decolorize recalcitrant dyes without addition of redox mediators, high laccase activities, high thermostability and efficient decolorization at 70 °C and pH 7.0, even in the presence of high concentrations of heavy metals (100 mM Cu⁺², Pb⁺² or Cd⁺²) or in a synthetic dyebath, T. trogii culture fluids could be effectively used to decolorize synthetic dyes from effluents.     

Evidence on manganese peroxidase and tyrosinase expression during decolorization of textile industry dyes by Trichosporon akiyoshidainum

Textile dyes are engineered to be resistant to environmental conditions. During recent years the treatment of textile dye effluents has been the focus of significant research because of the potentially low cost of the process. Mechanisms of biological textile dye decolorization depend greatly on the chemical structure of the dye and the microorganisms used. While basidiomycetous filamentous fungi are well recognized for dye decolorization through ligninolytic enzymes, reports on textile dye decolorization mechanisms of basidiomycetous yeasts have been scarce. Decolorization of several textile dyes by Trichosporon akiyoshidainum occurs during the first 12 h of cultivation. This fast decolorization process could not be solely related to siderophore production or dye sorption to biomass; it was shown to be a co-metabolic process. T. akiyoshidainum could use glucose, sucrose, and maltose as alternative carbon sources, and urea as an alternative nitrogen source with similar decolorization rates. The activity of two enzymes, manganese peroxidase and tyrosinase, were induced by the presence of dyes in the culture media, pointing to their potential role during the decolorization process. Manganese peroxidase titers reached 666 U l⁻¹ to 10538 U l⁻¹, while tyrosinase titers ranged between 84 U l⁻¹ and 786 U l⁻¹, depending on the dye tested. The present work provides a useful background to propose new eco-friendly alternatives for wastewater treatment in textile dying industries.     

Dynamics of microbial community for X-3B wastewater decolorization coping with high-salt and metal ions conditions

In this study, decolorization of Reactive Brilliant Red X-3B wastewater by the biological process coping with high salinity and metal ions conditions was investigated, and 16S rDNA based fingerprint technique was used to investigate microbial population dynamics. Results of sequencing batch tests showed that the microbial community could keep efficient with high concentration of dye (1100 mg L⁻¹), salt (150 g L⁻¹ NaCl) and some metal ions such as Mg²⁺, Ca²⁺ (1–10 mmol L⁻¹) and Pb²⁺ (1 mmol L⁻¹). 16S rDNA-based molecular analysis techniques demonstrated that the microbial community shifted during the acclimatization process affected by salt or metal ions. Some stains similar to Bacillus, Sedimentibacter, Pseudomonas, Clostridiales, Streptomyces and some uncultured clones acted for the dynamic succession, supposed as potential decolorization bacteria. This study provided insights on the decolorization capability and the population dynamic shifts during decolorization process of azo dye wastewater coping with salt and metal ions.     

A batch decolorization and kinetic study of Reactive Black 5 by a bacterial strain Enterobacter sp. GY-1

An Enterobacter strain (GY-1) with high activity of decolorization of Reactive Black 5 (RB 5) was isolated from textile wastewater treating sludge. The kinetic characteristics of dye decolorization by the strain GY-1 were determined quantitatively using the diazo dye, RB 5. Effects of different operation parameters (inoculum size, pH, temperature and salinity) and various electron donors on decolorization of the azo dye by GY-1 were systematically investigated to reveal the primary factors that determine the performance of the azo dye decolorization. The decolorization of RB 5 was attributed to extracellular enzymes. A kinetic model was established giving the dependence of decolorization rate on cell mass concentration (first order). Decolorization rate increased with increasing temperature from 20 to 35 °C, which can be predicted by Arrhenius equation with the activation energy (E_a) of 8.50 kcal mol⁻¹ and the frequency factor (A₀) of 6.28 × 10⁷ mg l g MLSS⁻¹ h⁻¹. Michaelis-Menten kinetics and Eadie-Hofstee plot were used to determine V_max, 1.05 mg l⁻¹ h⁻¹ and K_m, 24.06 mg l⁻¹.     

Assessment of olive-mill wastewater as a growth medium for lipase production by Candida cylindracea in bench-top reactor

Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l⁻¹ NH₄Cl and 3 g l⁻¹ olive oil led to an enzyme activity of about 10 U ml⁻¹. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml⁻¹) when the pH was held constant at 6.5, significantly increased (18.7 U ml⁻¹) with uncontrolled pH and was maximum (20.4 U ml⁻¹) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml⁻¹) and mean volumetric productivity (i.e., 123.5 U l⁻¹ h⁻¹).     

Feasibility of biogas production from anaerobic co-digestion of herbal-extraction residues with swine manure

The objective of this work was to examine the feasibility of biogas production from the anaerobic co-digestion of herbal-extraction residues with swine manure. Batch and semi-continuous experiments were carried out under mesophilic anaerobic conditions. Batch experiments revealed that the highest specific biogas yield was 294 mL CH₄ g⁻¹ volatile solids added, obtained at 50% of herbal-extraction residues and 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Specific methane yield from swine manure alone was 207 mL CH₄ g⁻¹ volatile solid added d⁻¹ at 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Furthermore, specific methane yields were 162, 180 and 220 mL CH₄ g⁻¹ volatile solids added d⁻¹ for the reactors co-digesting mixtures with 10%, 25% and 50% herbal-extraction residues, respectively. These results suggested that biogas production could be enhanced efficiently by the anaerobic co-digestion of herbal-extraction residues with swine manure.     

A spectrophotometric transesterification-based assay for lipases in organic solvent

A new method to evaluate lipase activities in nonaqueous conditions using vinyl ester absorbance at ultraviolet (UV) wavelengths is described. The model reaction is the transesterification between vinyl stearate and pentanol in hexane at 30 °C or in decane at 50 °C. The conversion of vinyl stearate into pentyl stearate is monitored through decreasing UV absorbance at 200 nm. Six commercial lipases were tested with this method, and results were compared with gas chromatography (GC) quantification and a classical spectrophotometric method using p-nitrophenyl palmitate. Results from the new spectrophotometric assay are similar both to results from GC quantification (R² = 0.999) and to results from p-nitrophenyl palmitate (R² = 0.989). The proposed method is able to evaluate both high activity from immobilized lipases such as immobilized Candida antarctica B lipase (3060 ± 350 U g⁻¹) and low activity from crude enzymatic extracts such as Carica papaya dried latex (0.1 ± 0.04 U g⁻¹). The method has also been used to measure kinetic parameters of C. antarctica B lipase for vinyl stearate and the correlation between its synthesis activity and its concentration. The method has also proved to be effective in studying the acyl selectivity of a lipase by comparing its activities with increasing chain lengths of vinyl esters.     

A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria

Metagenomic resources representing ruminal bacteria were screened for novel exocellulases using a robotic, high-throughput screening system, the novel CelEx-BR12 gene was identified and the predicted CelEx-BR12 protein was characterized. The CelEx-BR12 gene had an open reading frame (ORF) of 1140 base pairs that encoded a 380-amino-acid-protein with a predicted molecular mass of 41.8 kDa. The amino acid sequence was 83% identical to that of a family 5 glycosyl hydrolase from Prevotella ruminicola 23. Codon-optimized CelEx-BR12 was overexpressed in Escherichia coli and purified using Ni–NTA affinity chromatography. The Michaelis–Menten constant (K_m value) and maximal reaction velocity (V_max values) for exocellulase activity were 12.92 μM and 1.55 × 10⁻⁴ μmol min⁻¹, respectively, and the enzyme was optimally active at pH 5.0 and 37 °C. Multifunctional activities were observed against fluorogenic and natural glycosides, such as 4-methylumbelliferyl-β-d-cellobioside (0.3 U mg⁻¹), CMC (105.9 U mg⁻¹), birch wood xylan (132.3 U mg⁻¹), oat spelt xylan (67.9 U mg⁻¹), and 2-hydroxyethyl-cellulose (26.3 U mg⁻¹). Based on these findings, we believe that CelEx-BR12 is an efficient multifunctional enzyme as endocellulase/exocellulase/xylanase activities that may prove useful for biotechnological applications.     

Synergistic production of L-arabinose from arabinan by the combined use of thermostable endo- and exo-arabinanases from Caldicellulosiruptor saccharolyticus

The optimum conditions for the production of l-arabinose from debranched arabinan were determined to be pH 6.5, 75 °C, 20 g l⁻¹ debranched arabinan, 42 U ml⁻¹ endo-1,5-α-l-arabinanase, and 14 U ml⁻¹ α-l-arabinofuranosidase from Caldicellulosiruptor saccharolyticus and the conditions for sugar beet arabinan were pH 6.0, 75 °C, 20 g l⁻¹ sugar beet arabinan, 3 U ml⁻¹ endo-1,5-α-l-arabinanase, and 24 U ml⁻¹ α-l-arabinofuranosidase. Under the optimum conditions, 16 g l⁻¹l-arabinose was obtained from 20 g l⁻¹ debranched arabinan or sugar beet arabinan after 120 min, with a hydrolysis yield of 80% and a productivity of 8 g l⁻¹ h⁻¹. This is the first reported trial for the production of l-arabinose from the hemicellulose arabinan by the combined use of endo- and exo-arabinanases.     

Does scope for growth change as a result of salinity stress in the amphipod Gammarus oceanicus?

Physiological performance (feeding, metabolism, growth and excretion) across a broad range of salinity (5-30 psu) were determined for the benthic amphipod Gammarus oceanicus, a species of marine origin inhabiting brackish waters of the southern Baltic Sea. Feeding rates decreased with increasing salinity, whereas the nutritive absorption efficiency increased. Faeces production and ammonia excretion rates decreased strongly from the lowest to the highest salinity by 60% and 58%, respectively. Increasing salinity was accompanied by a reduction in the metabolic rate from 438 J g^− 1 dry wt d^− 1 (5.1 mW g^− 1) at 5 psu to 245 J g^− 1 (2.8 mW g^− 1) at 30 psu. Individuals were able to maintain a positive energy balance at all experimental salinities. The greatest values for scope for growth were recorded at the environmental salinity (7 psu) with a mean of 769 J g^− 1 dry wt d^− 1 (8.7 mW g^− 1).     

Potential for methane production from anaerobic co-digestion of swine manure with winery wastewater

This work examines the methane production potential for the anaerobic co-digestion of swine manure (SM) with winery wastewater (WW). Batch and semi-continuous experiments were carried out under mesophilic conditions. Batch experiments revealed that the highest specific methane yield was 348 mL CH₄ g⁻¹ COD added, obtained at 85.4% of WW and 0.7 g COD g⁻¹ VS. Specific methane yield from SM alone was 27 mL CH₄ g⁻¹ COD added d⁻¹. Furthermore, specific methane yields were 49, 87 and 107 mL CH₄ g⁻¹ COD added d⁻¹ for the reactors co-digesting mixtures with 10% WW, 25% WW and 40% WW, respectively. Co-digestion with 40% WW improved the removal efficiencies up to 52% (TCOD), 132% (SCOD) and 61% (VSS) compared to SM alone. These results suggest that methane can be produced very efficiently by the co-digestion of swine manure with winery wastewater.     

Influence of exogenous salicylic acid on flavonolignans and lipoxygenase activity in the hairy root cultures of Silybum marianum

Silymarin is one of the most potent antioxidant so far developed from plant sources used as hepatoprotectants. Influence of different concentrations (0, 1, 2, 4, 6 and 8 mg/50 ml culture) and exposure time (24, 48, 72, 96 and 120 h) of salicylic acid on lipoxygenase activity, linoleic acid content, growth and production of silymarin in hairy root cultures of S. marianum were investigated. Detection and identification of flavonolignans was carried out by high performance liquid chromatograph method. Salicylic acid enhanced silymarin production (1.89 mg g⁻¹ DW). The optimal feeding condition was the addition of salicylic acid (6 mg/50 ml culture) after 24 h in which the silymarin content was 2.42 times higher than the control (0.78 mg g⁻¹ DW). The content of silybin, isosilybin, silychristin, silydianin and taxifolin were 0.703, 0.017, 0.289, 0.02 and 0.863 mg g⁻¹ DW respectively in these samples, while in non-treated hairy roots were 0.027, 0.046, 0.23, 0.022 and 0.453 respectively. Lipoxygenase activity also affected by elicitation. lipoxygenase activity increased 24 h after treatment by ∼1.57- fold (0.21 Δ OD₂₃₄/mg protein min⁻¹). Upon elicitation with salicylic acid, linoleic acid content of hairy roots (38.26 mg g⁻¹ DW) were also elevated after 24 h, in which the linoleic acid content was 2.37 times higher than the control (16.1 mg g⁻¹ DW). It is feasible that elicitation with salicylic acid regulates the jasmonate pathway, which in turn mediates the elicitor-induced accumulation of silymarin.     

Conformational stability of α-amylase from malted sorghum (Sorghum bicolor): Reversible unfolding by denaturants

α-Amylase from Sorghum bicolor, is reversibly unfolded by chemical denaturants at pH 7.0 in 50 mM Hepes containing 13.6 mM calcium and 15 mM DTT. The isothermal equilibrium unfolding at 27 °C is characterized by two state transition with ΔG (H₂O) of 16.5 kJ mol⁻¹ and 22 kJ mol⁻¹, respectively, at pH 4.8 and pH 7.0 for GuHCl and ΔG (H₂O) of 25.2 kJ mol⁻¹ at pH 4.8 for urea. The conformational stability indicators such as the change in excess heat capacity (ΔC_p), the unfolding enthalpy (H_g) and the temperature at ΔG = 0 (T_g) are 17.9 ± 0.7 kJ mol⁻¹ K⁻¹, 501.2 ± 18.2 kJ mol⁻¹ and 337.3 ± 6.9 K at pH 4.8 and 14.3 ± 0.5 kJ mol⁻¹ K⁻¹, 509.3 ± 21.7 kJ mol⁻¹ and 345.4 ± 4.8 K at pH 7.0, respectively. The reactivity of the conserved cysteine residues, during unfolding, indicates that unfolding starts from the ‘B’ domain of the enzyme. The oxidation of cysteine residues, during unfolding, can be prevented by the addition of DTT. The conserved cysteine residues are essential for enzyme activity but not for the secondary and tertiary fold acquired during refolding of the denatured enzyme. The pH dependent stability described by ΔG (H₂O) and the effect of salt on urea induced unfolding confirm the role of electrostatic interactions in enzyme stability.     

Comparison of stirred tank and airlift bioreactors in the production of polygalacturonases by Aspergillus oryzae

The production of endo and exo-polygalacturonase (PG) by Aspergillus oryzae IPT 301 was studied in a stirred tank bioreactor (STR) and an internal circulation airlift bioreactor. Using a factorial experimental design, a soluble culture medium was defined which allowed the production of exo- and endo-PG comparable to that obtained in a medium containing suspended wheat bran. The soluble medium was used in tests to compare the production of these enzymes in the STR and airlift bioreactor. In these tests, after 96 h, maximum enzymatic activity values achieved for exo- and endo-PG were 65.2 units (U) per mL and 91.3 U mL⁻¹, in the STR, with similar activity values of 60.6 U mL⁻¹ and 86.2 U mL⁻¹, respectively, being achieved in the airlift bioreactor. The airlift bioreactor also showed satisfactory results regarding the oxygen transfer rate in this process, indicating its potential to be used in an eventual larger scale production of exo- and endo-PG, with lower costs for both installation and operation.     

Evaluation of selenite bioremoval from liquid culture by Enterococcus species

The genus Enterococcus belong to the genera of bacteria that produce lactic acid and can confer health benefits to living organisms. Selenium (Se) is an essential micronutrient for humans and animals. Thirty-six Enterococcus species isolated from dairy products were screened for Se(IV) sorption capacity for use as a probiotics in animal nutrition. Several isolates grew luxuriantly and significantly removed Se(IV) from Se(IV) amended medium. Two isolates, LAB 14 and LAB 18, identified by 16S rRNA gene sequence analysis as Enterococcus faecalis (98% nucleotide sequence similarity) and Enterococcus faecium (97% nucleotide sequence similarity), respectively, were selected for further studies. The two isolates grew optimally and removed selenium at initial pH 7.0. Optimum removal of Se(IV) from the medium was recorded at 25 °C. Time course studies showed that after 8 h of incubation LAB 14 and LAB 18 cultures displayed the highest biomass production and Se(IV) bioremoval and most selenite in culture depleted in 24 h. At initial concentrations of 10 mg L⁻¹ and 60 mg L⁻¹, E. faecium (LAB 18) removed 9.91 mg L⁻¹ and 59.70 mg L⁻¹, respectively after 24 h. Similar Se(IV) bioremoval capacity was recorded with E. faecalis (LAB 14). Substantial amount of Se was detected in biomass of E. faecium (0.4599 mg g⁻¹ of dry weight) and E. faecalis (0.4759 mg g⁻¹ of dry weight). The significant uptake and transformation of Se(IV) by the Enterococcus species observed in this study suggest that they can be used to deliver dietary Se through feed augmentation with Se(IV)-enriched Enterococcus biomass.     

Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis

Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD₅) (from an inlet of 420 to 1000 mg L⁻¹) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L⁻¹), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P < 0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha⁻¹ d⁻¹ and 529 kg BOD₅ ha⁻¹ d⁻¹ were achieved for a loading ranging from 242 to 1925 kg COD ha⁻¹ d⁻¹ and from 126 to 900 kg BOD₅ ha⁻¹ d⁻¹. Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation.     

Copper toxicity in gibel carp Carassius auratus gibelio: Importance of sodium and glycogen

In general, copper is primarily an osmoregulatory toxicant to fish and Cu toxicity is thought to be related to the rate of sodium loss. Looking at a previous research it is striking that gibel carp, Carassius auratus gibelio, do not seem so susceptible to the first ionoregulatory shock phase of Cu exposure, but rather build up physiological disturbances slowly until mortality occurs. Since it was noted that gibel carp experience severe hypoxia under Cu exposure, we hypothesised that, besides the Na loss, the slow depletion of liver glycogen stores contributed equally to the collapse of physiological integrity. It is clear from our results that glycogen stores are being depleted in Cu exposed fish and that dead fish suffered more extensive glycogen losses compared to surviving fish, with liver glycogen levels of 125 ± 8 mg g⁻¹ in dead fish compared to 230 ± 13 mg g⁻¹ in surviving fish. However, changes in liver glycogen did not contribute significantly to mortality, while changes in whole body sodium and the rate of sodium loss did. Whole body Na levels dropped from 1111 ± 48 μg g⁻¹ dry weight in control fish to 850 ± 54 μg g⁻¹ in surviving fish and to 607 ± 24 μg g^− 1 in fish that had died resulting in Na loss rates of 1.25 ± 0.22 μg g^− 1 h^− 1 and 3.39 ± 0.19 μg g^− 1 h^− 1 in surviving and dead fish respectively. Our results support the finding that the rate of Na loss largely determines Cu toxicity in fish, even in resistant species.     

Decomposition of Phragmites australis rhizome in a shallow lake

Decomposition of Phragmites australis (Cav. Trin ex Steudel) rhizome was studied at Lake Fert?/Neusiedler See using the litter bag technique. Samples were analysed for rhizome dry mass, fibre (cellulose, hemicellulose, lignin) and nutrient content (C, N, P and S), litter-associated fungal biomass, potential microbial respiration (electron transport activity: ETS) and cellulolitic bacteria. The mass loss of decomposing rhizome was rapid in the initial period and only 13.6% of the dry mass remained at the end of the experiment during 953 days. Substantial quantities of C, N, S and P were lost during 99 days; only 18% C, 19% N, 14% S and 6.4% of the P remained after 953 days. Hemicellulose degraded more rapidly than the other fibres whilst the lignin had the slowest rate of decomposition. Bacteria were found to be the primary colonizers of plant detritus, which was followed by fungal growth. An antagonistic relationship was observed between bacteria and fungi. Fungal biomass as determined by ergosterol concentrations ranged between 4.1 and 420 μg g⁻¹ and peaked every year in September. The number of cellulolitic bacteria varied from 0 to 22 MPN g⁻¹ with higher values in summer. The ETS-activity ranged between 0.1 and 1.6 mg O₂ g⁻¹ h⁻¹. The changes in ETS-activity varied almost in parallel with the in situ temperature of the lake water.     

Bicarbonate use in three aquatic plants

Our study aimed to test the ability of aquatic plants to use bicarbonate when acclimated to three different bicarbonate concentrations. To this end, we performed experiments with the three species Ceratophyllum demersum, Egeria densa, Lagarosiphon major to determine photosynthetic rates under varying bicarbonate concentrations. We measured bicarbonate use efficiency, photosynthetic performance and respiration. For all species, our results revealed that photosynthetic rates were highest in replicates grown at low alkalinity. Thus, E. densa had approx. five times higher rates at low (264 ± 15 μmol O₂ g⁻¹ DW h⁻¹) than at high alkalinity (50 ± 27 μmol O₂ g⁻¹ DW h⁻¹), C. demersum had three times higher rates (336 ± 95 and 120 ± 31 μmol O₂ g⁻¹ DW h⁻¹), and L. major doubled its rates at low alkalinity (634 ± 114 and 322 ± 119 μmol O₂ g⁻¹ DW h⁻¹). Similar results were obtained for bicarbonate use efficiency by E. densa (136 ± 44 and 43 ± 10 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹) and L. major (244 ± 29 and 82 ± 24 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹). As to C. demersum, efficiency was high but unaffected by alkalinity, indicating high adaptation ability to varied alkalinities. A pH drift experiment supported these results. Overall, our results suggest that the three globally widespread worldwide species of our study adapt to low inorganic carbon availability by increasing their efficiency of bicarbonate use.     

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 μg PCP g⁻¹ VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l⁻¹ day⁻¹ for R1, and from 0.06 to 4.15 mg PCP l⁻¹ day⁻¹ for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m⁻³ day⁻¹ at hydraulic retention times (HRT) of 24 h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.