A study was made of the metabolic and co-metabolic intermediates of 2- and 3-chlorobenzoate, 2,3- and 3,5-dichlorobenzoate to elucidate the mechanism(s) involved in the negative effects observed on the growth of a chlorobenzoate-degrading microbial consortium in the presence of mixed chlorobenzoates. 2-Chloromuconate accumulated as the end-product in the cultural broths of the microbial consortium during growth on 2-chlorobenzoate; the same 2-chloromuconate was identified in the reaction mixtures of resting cells pregrown on 2-chlorobenzoate and exposed to 3-chloro- and 2,3-dichlorobenzoate, while in similar experiments 1,2-dihydroxy-3,5-dichloro-cyclohexa-3,5-dienoate was detected as dead-end product of 3,5-dichlorobenzoate co-metabolism. These results suggest an initial degradative attack by 2-chlorobenzoate induced dioxygenase(s). The role of 3,5-dichlorobenzoate as an antagonist of 2-chlorobenzoate degradation was also studied: in the presence of mixed 2-chloro- and 3,5-dichlorobenzoate, the 3,5-dichlorobenzoate preferential uptake by the resting cells of the chlorobenzoate-degrading consortium was observed. 2-Chlorobenzoate entered the cells only after the complete removal of the co-substrate. In growing cells experiments, the addition of 1,2-dihydroxy-3,5-dichloro-cyclohexa-3,5-dienoate, the 3,5-dichlorobenzoate co-metabolite, to 2-chlorobenzoate exerted the same antagonistic effect of the parent compound, inhibiting both the microbial growth and the degradative process. These data are discussed, allowing us to attribute the inhibitory effects observed to a substrate/co-substrate competition, though other additional causes may not be totally excluded.
A selected mixed culture and a strain of Alcaligenes eutrophus TCP were able to totally degrade 2,4,6‐TCP with stoichiometric release of Cl−. In cultures of Alc. eutrophus TCP, a dioxygenated dichlorinated metabolite was detected after 48 h of incubation. Experiments conducted with soil microcosms gave evidence that : the degradative process had a biotic nature and was accompanied by microbial growth ; the soil used presented an intrinsic degradative capacity versus 2,4,6‐TCP ; the specialized organism used as inoculum was effective in degrading 2,4,6‐TCP in a short time. These results could be utilized for the adoption of appropriate remediation techniques for contaminated soil.
Two soils of different contamination history were tested in slurry for their self-remediability towards mono-, di- and trisubstituted chlorophenols. The landfill soil showed poor ability in removing the compounds. Instead, the soil from the golf course, treated for many years with a 2,4,6-trichlorophenol derivative (Prochloraz), remediated different concentrations of the same 2,4,6TCP, 2,4-dichlorophenol and monochlorophenol isomers, singly and in mixtures, at varying degradation rates. Ralstonia eutropha TCP, a specialised microorganism capable of degrading 2,4,6TCP, proved highly efficient in removing the compound from both tested soils. The same microbial inoculum allowed total removal of the ternary mixture of monochlorophenol isomers from the golf course soil, but it did not accelerate the removal of the same compounds when singly supplied. The addition of phenol as a degradable analogue was more effective in co-metabolically removing not only the single monochlorophenols, but also their mixtures, the removal occurring faster and independently of the presence of the microbial inoculum. From the golf course soil, a microorganism, phenotypically and genetically identical to R. eutropha TCP, was isolated and classified as R. eutropha TCP II.
The degradation of mono‐ and dichlorobenzoates was followed in soil suspensions. The indigenous microflora of the soil used was able to degrade only 4‐chlorobenzoic acid (4CB) and 2‐chlorobenzoic acid (2CB) either added alone or present in a mixture. When the soil was inoculated with an Alcaligenes denitrificans CB strain capable of degrading 4CB, 4CB degradation was highly accelerated. The presence in the soil suspensions of other chlorobenzoates (CBs) together with 4CB affected the degradation rate of the latter compound: 2CB and 2,6‐dichlorobenzoic acid (2,6‐DCB) retarded the degradative process, while 3,4‐ or 2,3‐dichlorobenzoic acids (3,4‐, 2,3‐DCB) totally inhibited 4CB degradation. The potential mechanisms involved in these interactions are discussed.
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