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Effects of xylene isomers on operant responding and motor performance in mice.

Xylene, a widely used industrial solvent, is a mixture of the ortho-, meta-, and paraisomers. In this study we examined the effects of each individual isomer, as well as a commercial-grade mixture of xylenes, on two behavioral measures: (1) operant performance of 15 mice trained to lever-press under a DRL (differential reinforcement of low rates) 10-sec schedule, and (2) motor performance of mice on an inverted screen test. The 15-min operant sessions immediately followed 30-min exposures to solvent vapors (500 to 7000 ppm), or air, in static inhalation chambers. All mice were exposed to each isomer in a counterbalanced order, and the mixture was given last in all cases. Ortho-, meta-, para-, and mixed xylenes produced similar biphasic effects on response rates, and concentration-dependent decreases in reinforcement rates. The lowest significantly effective concentration for each isomer on any variable was 1400 ppm, where increases in response rates occurred. Half-maximal response rate decreases were produced by 5179 ppm (ortho-xylene) to 6176 ppm (meta-xylene). The temporal distribution of responses was only moderately disrupted, even at high concentrations. In other groups of mice, motor coordination was also disrupted by the xylenes in a concentration-dependent manner. Half-maximal effective concentrations were 2676 ppm (para-xylene) to 3790 ppm (meta-xylene). Minimally effective concentrations were 2000 to 3000 ppm, higher than those seen in the operant studies. Xylene produced pronounced behavioral actions following acute exposure and no substantial differences in overall effects, and only slight potency differences, were obtained between the individual isomers or the commercial mixture.

Animals↗

Conditioned flavor aversion induced by inhaled p-xylene in rats.

Xylenes have the potential for widespread public exposure, yet their neurotoxic properties are poorly understood. The conditioned flavor aversion (CFA) paradigm provides a sensitive behavioral assay for the aversiveness of many drugs and toxic chemicals. Male Long-Evans rats were placed on a restricted water schedule (30 min/day) 1 week after arrival in the laboratory at 40 days of age. Ten days later, all rats received 0.1% saccharin in place of water, and then were exposed immediately either to filtered air or to 50, 100, 200, 400, 800 or 1,600 ppm p-xylene for 4 hr, or to air or 400 ppm p-xylene for 0.5, 1, 2, 4 or 8 hr. The restricted water schedule remained in effect for the next 72 hr, at which time the rats were given a choice between saccharin and water. Inhalation of all concentrations of p-xylene reduced relative saccharin intake, with maximal aversion at 800 and 1,600 ppm. The aversion produced by 400 ppm p-xylene was maximal at exposures of 2 or more hr, with shorter exposures producing intermediate effects. Total fluid intake was unaffected by p-xylene exposure with the exception of a slight (nonsignificant) decrease in consumption following 1,600 ppm. Exposure to saccharin 24 hr prior to p-xylene produce no aversion, indicating that the reduction in saccharin intake required close temporal pairing of saccharin and p-xylene. Thus inhaled p-xylene at a concentration of 1/2 its TLV (= 100 ppm) caused a significant, learned change in rats' normal consumption of saccharin-flavored water, without disrupting total fluid consumption.

Administration, Inhalation↗

The use of a solid adsorber resin for enrichment of bacteria with toxic substrates and to identify metabolites: degradation of naphthalene, O-, and m-xylene by sulfate-reducing bacteria.

Anaerobic sulfate-reducing bacteria were enriched from contaminated aquifer samples with naphthalene, o-, and m-xylene as sole carbon and energy source in the presence of Amberlite-XAD7, a solid adsorber resin. XAD7 served as a substrate reservoir maintaining a constantly low substrate concentration in the culture medium. In equilibration experiments with XAD7, the aromatic hydrocarbons needed up to 5 days to achieve equilibrium between the water and the XAD7 phase. The equilibrium concentration was directly correlated with the amount of added substrate and XAD7. In the enrichments presented here, XAD7 and aromatic hydrocarbons were adjusted to maintain substrate concentrations of 100 microM m-, or o-xylene, or 50 microM naphthalene. After five subsequent transfers, the three cultures were able to grow with higher substrate concentrations in the absence of XAD7 although they grew best with lower hydrocarbon concentrations. Two new xylene-degrading cultures were obtained that could not utilise toluene as carbon source. O-xylene was degraded anaerobically by a culture, which could also oxidise m-xylene but not p-xylene. Eighty-three percent of the electrons from o-xylene oxidation were recovered in the produced sulfide, indicating a complete oxidation to CO2. Another sulfate-reducing enrichment culture oxidised m-xylene completely to CO2 but not o-, or p-xylene. A naphthalene-degrading sulfate-reducing enrichment culture oxidised naphthalene completely to CO2. Metabolites of naphthalene degradation were recovered from the XAD7 phase and subjected to GC/MS analysis. Besides the metabolites 2-naphthoic acid and decahydro-2-naphthoic acid which were identified by the mass spectrum and coelution with chemically synthesised reference compounds, the reduced 2-naphthoic acid derivatives 5,6,7,8-tetrahydro-2-naphthoic acid and octahydro-2-naphthoic acid were tentatively identified by their mass spectra. Cultivation of bacterial cultures in the presence of XAD7 and subsequent derivatisation and extraction of metabolites directly from the solid XAD7 resin provides a new method for the isolation of sensitive bacteria and identification of metabolites.

Acrylic Resins↗

Xylene-free method for histological preparation: a multicentre evaluation.

The backbone of daily pathological diagnostic work is the paraffin section. Paraffin sections are still prepared by methods largely unchanged for over 150 years. A xylene-free method has been developed that excludes xylene, not only as the intermediate step before the paraffin baths, but also for deparaffinizing of the cut sections, which also eliminates the need for rehydration and dehydration for the staining and mounting steps. Elimination of xylene from tissue processing cuts costs, saves time, and improves the laboratory environment. Experience with xylene-free sections since 1995 at the Vrinnevi Hospital is favorable. Our opinion is that the xylene-free sections are equivalent to conventionally processed sections. To test this hypothesis, nine pathologists from three hospitals participated in an evaluation trial. Paired tissue blocks from 10 consecutively submitted samples each of breast, intestine, and skin were processed by either the xylene-free or the conventional method. Sections from each block were deparaffinized and stained with hematoxylin-eosin (H&E), with periodic acid-Schiff (PAS), and with van Gieson's method. A randomized mix of 180 sections (10 samples x 3 tissues x 3 stains x 2) gave 90 matched pairs. Each section was blindly examined and scored by nine pathologists to give 810 paired observations for statistical evaluation. The xylene-free sections were ranked as good as or better than their conventional counterparts in 74% of the comparisons, and poorer in 26%. The major discriminating factor was the staining method. H&E and PAS sections were equivalent. The xylene-free van Gieson sections, cut from the same blocks and randomly assigned to this stain, tended to be downgraded. This could be traced to a faulty stain solution used for this batch. The overall results have demonstrated professional acceptance for the xylene-free method of processing histological sections.

Breast↗

Evaluation of the dermal bioavailability of aqueous xylene in F344 rats and human volunteers.

Xylene is a clear, colorless liquid used as a solvent in the printing, rubber, and leather industries and is commonly found in paint thinners, paints, varnishes, and adhesives. Although humans are most likely to be exposed to xylene via inhalation, xylene is also found in well and surface water. Therefore, an assessment of the dermal contribution to total xylene uptake is useful for understanding human exposures. To evaluate the significance of these exposures, the dermal absorption of o-xylene was assessed in F344 male rats and human volunteers using a combination of real-time exhaled breath analysis and physiologically based pharmacokinetic (PBPK) modeling. Animals were exposed to o-xylene dermally. Immediately following the initiation of exposure, individual animals were placed in a glass off-gassing chamber and exhaled breath was monitored. Human volunteers participating in the study placed both legs into a stainless steel hydrotherapy tub containing an initial concentration of approximately 500 microg/L o-xylene. Exhaled breath was continually analyzed from each volunteer before, during, and after exposure to track absorption and subsequent elimination of the compound in real time. In both animal and human studies, a PBPK model was used to estimate the dermal permeability coefficient (K(p)) to describe each set of exhaled breath data. Rat skin was found to be approximately 12 times more permeable to aqueous o-xylene than human skin. The estimated human and rat aqueous o-xylene K(p) values were 0.005 +/- 0.001 cm/h and 0.058 +/- 0.009 cm/h, respectively.

Absorption↗

Teratogenicity of a commercial xylene mixture in the mouse.

Pregnant outbred albino (CD-1) mice received (gavage, three times a day in cottonseed oil) a xylene mixture (60.2% m-xylene, 9.1% o-xylene, 13.6% p-xylene, and 17.0% ethyl benzene) on d 6-15 of gestation (d 1 being the day vaginal plugs were observed). The mice were killed on d 18, the general and reproductive health of the dams evaluated, and the fetuses examined and processed to characterize external, visceral, and skeletal malformation. At 3.6 ml/kg.d, xylene killed 12 of 38 dams and caused a significantly (p less than 0.05) smaller average weight gain during pregnancy than did the vehicle (cottonseed oil). Fetuses from dams treated with xylene at 2.4 ml/kg.d and higher doses had average fetal weights significantly lower than that of the control fetuses. However, the percent of resorptions for xylene was significantly greater than for the control only at 3.6 ml/kg-d. At 2.4, 3.0, and 3.6 ml/kg-d xylene produced a significantly (p less than 0.01) greater average percent of malformed fetuses than did the control. Cleft palate was the major malformation at all three doses. When bilateral (multiple) wavy ribs were counted as a malformation, the average percent of malformed fetuses increased from 7.8 to 10.5 at 3.0 ml/kg.d and from 9.1 to 13.4 at 3.6 ml/kg.d. It is concluded that xylene (mixed isomers) is teratogenic to the CD-1 mouse at 2.4 and 3.0 ml/kg.d, doses approaching lethal levels.

Animals↗

Cloning of the genes for and characterization of the early stages of toluene and o-xylene catabolism in Pseudomonas stutzeri OX1.

In order to study the toluene and o-xylene catabolic genes of Pseudomonas stutzeri OX1, a genomic library was constructed. A 28-kb EcoRI restriction endonuclease DNA fragment, cloned into the vector plasmid pLAFR1 and designated pFB3401, permitted Pseudomonas putida PaW340 to convert toluene and o-xylene into the corresponding meta-ring fission products. Physical and functional endonuclease restriction maps have been derived from the cloned DNA fragment. Further subcloning into and deletion analysis in the Escherichia coli vector pGEM-3Z allowed the genes for the conversion of toluene or o-xylene into the corresponding catechols to be mapped within a 6-kb region of the pFB3401 insert and their direction of transcription to be determined. Following exposure to toluene, E. coli cells carrying this 6-kb region produce a mixture of o-cresol, m-cresol, and p-cresol, which are further converted to 3-methylcatechol and 4-methylcatechol. Similarly, a mixture of 2,3-dimethylphenol and 3,4-dimethylphenol, further converted into dimethylcatechols, was detected after exposure to o-xylene. The enzyme involved in the first step of toluene and o-xylene degradation exhibited a broad substrate specificity, being able to oxidize also benzene, ethylbenzene, m-xylene, p-xylene, styrene, and naphthalene. Deletions of the 6-kb region which affect the ability to convert toluene or o-xylene into the corresponding methylphenols compromise also their further oxidation to methylcatechols. This suggests that a single enzyme system could be involved in both steps of the early stages of toluene and o-xylene catabolism.

Benzene↗

Effects of acetone on the capacity of o-xylene and toluene to induce several forms of cytochrome P450 in rat liver.

The influence of acetone on the induction of P450 in lung, kidney and liver by toluene and xylene was studied in an attempt to estimate more precisely the range of P450 which are proposed to function in the activation of xylenes and toluene metabolites in the conditions of combined exposures. The addition of acetone potentiated the induction of CYP1A1/2 to 4 minutes ethoxyresorufin deethylation that from control animals after pretreatment with toluene. The level of this enzymatic activity (0.4 nmol/min/mgP) estimated in this study is consistent with the turnover number of the purified CYP1A1 (0.45 nmol/min/mgP). The assay activity results were confirmed by immunoblotting detection of the CYP1A1/2 levels. Oppositely, the addition of acetone in the inhalation protocol using o-xylene lead to a lower ethoxyresorufin deethylation activity as that in rat livers from rats pretreated with xylene alone. Pentoxyresorufin deethylation activity was enhanced strongly by toluene (3.5 times the controls) and by xylene (4 times the controls) and the addition of acetone potentiated the induction of CYP2B1 both after xylene and toluene. A 4 fold induction of CYP2E1 dependent chlorzoxazone hydroxylation after exposure to toluene and 2.5 fold after xylene corresponded to the enzyme levels of P45020E1 detected by immunoblotting. The addition of acetone consequently potentiated the induction effect of toluene and xylene and increased its levels more markedly in the case of xylene, which is a weaker inducer of CYP2E1.

Acetone↗

Ototoxicity in rats exposed to ethylbenzene and to two technical xylene vapours for 13 weeks.

Male Sprague-Dawley rats were exposed to ethylbenzene (200, 400, 600 and 800 ppm) and to two mixed xylenes (250, 500, 1,000 and 2,000 ppm total compounds) by inhalation, 6 h/day, 6 days/week for 13 weeks and sacrificed for morphological investigation 8 weeks after the end of exposure. Brainstem auditory-evoked responses were used to determine auditory thresholds at different frequencies. Ethylbenzene produced moderate to severe ototoxicity in rats exposed to the four concentrations studied. Increased thresholds were observed at 2, 4, 8 and 16 kHz in rats exposed to 400, 600 and 800 ppm ethylbenzene. Moderate to severe losses of outer hair cells of the organ of Corti occurred in animals exposed to the four concentrations studied. Exposure to both mixed xylenes produced ototoxicity characterized by increased auditory thresholds and losses of outer hair cells. Ototoxicity potentiation caused by ethylbenzene was observed. Depending on the mixed xylene studied and the area of the concentration-response curves taken into account, the concentrations of ethylbenzene in mixed xylenes necessary to cause a given ototoxicity were 1.7-2.8 times less than those of pure ethylbenzene. Given the high ototoxicity of ethylbenzene, the safety margin of less or equal to two (LOAEL/TWA) might be too small to protect workers from the potential risk of ototoxicity. Moreover, the enhanced ototoxicity of ethylbenzene and para-xylene observed in mixed xylenes should encourage the production of mixed xylenes with the lowest possible concentrations of ethylbenzene and para-xylene.

Air Pollutants, Occupational↗

Occupational chronic exposure to organic solvents XVII. Ambient and biological monitoring of workers exposed to xylenes.

OBJECTIVE: Ambient-air and biological monitoring of occupational xylene exposure were carried out on 2 groups of workers (13 and 10 men, respectively) exposed to a mixture of xylenes during the production of paints or during spraying. METHODS: Personal ambient-air monitoring was performed for one complete work shift. Blood and urine samples were collected directly at the end of the shift. Biological monitoring was based on the determination of the concentration of xylenes in blood and on the quantification of the sum of the three methylhippuric acids in urine. RESULTS: Average xylene ambient-air concentrations were 29 ppm (production) and 8 ppm (spraying), ranging from 5 to 58 ppm and from 3 to 21 ppm, respectively. The concentrations of xylenes in blood ranged from 63 to 715 microg/l and from 49 to 308 microg/l, with average values being 380 and 130 microg/l, respectively. Accordingly, the workers engaged in paint production also excreted more methylhippuric acids in their urine (average 1221 mg/l, range 194 2333 mg/l) than did the sprayers (average 485 mg/l, range 65-1633 mg/l). DISCUSSION: Our results as well as a literature review indicate that occupational xylene exposure on average barely exceeds the threshold limit value of 100 ppm as proposed by both American and German institutions. Biological monitoring based on the determination of xylenes in blood and of methylhippuric acids in urine provides sufficient sensitivity and specificity for occupational health surveillance. The results also confirm the current limit values (BAT values) proposed by the Deutsche Forschungsgemeinschaft for xylenes in blood (1500 microg/l) and methylhippuric acids in urine (2000 mg/l).

Adult↗

Effects of xylene exposure on the metabolism of antipyrine in vitro and in vivo in the rat.

Exposure of male rats to different concentrations of xylene for 3 days induced, in a dose-dependent way, the in vitro liver microsomal metabolism of antipyrine. The degree of induction was statistically significant at an exposure level of 250 ppm and maximal (2.5-fold increase) at 2000 ppm. This increase was of the same magnitude as after phenobarbital treatment. Female rats had a lower basal antipyrine metabolism than males, but exhibited a greater relative increase in antipyrine metabolism following xylene exposure. Cytochrome P-450 isozymes, purified from xylene- and phenobarbital-treated animals, were efficient catalysts of antipyrine metabolism, with turnover numbers of 33.3 and 21.1, respectively. A reduction in the half-life of antipyrine to 39% of preexposure values occurred after exposure of male rats to 1000 ppm of xylene for 3 days. Exposure to lower xylene levels did not produce significant alterations in antipyrine elimination half-life. In vitro, xylene was shown to be a non-competitive metabolic inhibitor of antipyrine. Experiments in vivo indicated that inhibition is not important at relatively low xylene exposure levels. It is concluded that induction of hepatic monooxygenases by xylene can be demonstrated, with antipyrine as a test drug, both in vitro and in vivo.

Animals↗

The contribution of sensory nerves to xylene-induced cystitis in rats.

The role of capsaicin-sensitive sensory nerves of the rat urinary bladder in xylene-induced cystitis was investigated. Instillation of xylene into the urinary bladder of female rats induced cystitis, e.g. detrusor hyperreflexia and increased vascular permeability. Detrusor hyperreflexia was also observed in rats desensitized to capsaicin as adults (50-125 mg/kg s.c., 4 days before) but only for a short period (1 h) after instillation. When a longer time lag (24 h) was allowed to elapse following instillation, reflex micturition was almost abolished. In rats desensitized to capsaicin as newborns (50 mg/kg s.c. on second day of life) reflex micturition was almost abolished and xylene (given 1 h before measurement) was ineffective. The xylene-induced plasma extravasation was greater in the bladder neck than in the dome. In the bladder neck the "early" response to xylene was reduced but not abolished in rats densensitized to capsaicin as adults or pretreated with compound 48/80 and was abolished in rats desensitized to capsaicin as newborns. The bladder content of substance P-like immunoreactivity decreased at various times following xylene instillation but this change occurred in parallel to the increase in bladder weight. These findings indicate that xylene-induced cystitis involves, at least in part, an irritation of capsaicin-sensitive sensory nerves in the bladder wall. The present results further suggest that xylene acts by stimulating at least two populations of sensory nerves which differ in their sensitivity towards capsaicin.

Animals↗

Behavioral effects of acute p-xylene inhalation in rats: autoshaping, motor activity, and reversal learning.

p-Xylene is a ubiquitous solvent and chemical precursor used in industry, gasoline, and household products. While the population at risk for exposure is thus quite large, little is known about its neurobehavioral effects. To evaluate the possibility that p-xylene affects cognitive behavior, male Long-Evans hooded rats inhaled p-xylene at concentrations of 0 or 1600 ppm, 4 hr per day for 1 to 5 days, and were evaluated after exposure on two learning tasks and a test of motor activity. Autoshaping was carried out across 5 successive days with p-xylene exposure in the morning followed by testing in the afternoon. For this test, the retraction of a single response lever on a variable-time 35-sec schedule was followed by delivery of a food pellet. When the force required to depress the lever was low (0.10 N), response acquisition was faster in animals having inhaled 1600 ppm p-xylene than in air-exposed controls. When the force was increased to 0.20 N, however, p-xylene-exposed rats acquired the response no faster than controls. In contrast, inhaled p-xylene at 1600 ppm suppressed response rates in an automaintained reversal learning paradigm without affecting reversal rate. Studies of motor activity showed that while vertically-directed activity was unaffected by p-xylene, horizontally-directed activity was increased by about 30% for the first 15 min of each daily 25-min test.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

Regioselective oxidation of xylene isomers by Rhodococcus sp. strain DK17.

Rhodococcus sp. strain DK17 is able to utilize a variety of monocyclic aromatic hydrocarbons, including benzene, phenol, toluene, and o-xylene, as growth substrates. Although DK17 is unable to grow on m- and p-xylene, this strain could transform these two xylene isomers to some extent after induction by o-xylene. The major accumulating compounds formed during the degradation of m- and p-xylene by DK17 were isolated by high-pressure liquid chromatography and identified by gas chromatography-mass spectrometric and (1)H nuclear magnetic resonance spectral techniques. Both xylene isomers were transformed to dihydroxylated compounds by what must be two successive hydroxylation events: m-xylene was converted to 2,4-dimethylresorcinol and p-xylene was converted to 2,5-dimethylhydroquinone. The rigorous structural identification of 2,4-dimethylresorcinol and 2,5-dimethylhydroquinone demonstrates that DK17 can perform distinct regioselective hydroxylations depending on the position of the substituent groups on the aromatic ring.

Hydroxylation↗

Xylene: its toxicity, measurement of exposure levels, absorption, metabolism and clearance.

Xylene is an aromatic hydrocarbon widely used in industry and medical technology as a solvent. Health and safety authorities in most countries, including Australia, recommend a threshold limit value (TLV) of 100 ppm in the working environment. Recently, the amount of the major metabolite of xylene, methylhippuric acid (MHA), in urine has been recommended as a better indicator of exposure. The American Conference of Governmental Industrial Hygienists has recommended an upper limit for this indicator, called a biological exposure index (BEI), of 2.0 g MHA/L urine (SG 1.016). Xylene vapour is absorbed rapidly from the lungs, and xylene liquid and vapour are absorbed slowly through the skin. Of the xylene absorbed, about 95% is metabolised in the liver to MHA and 70 to 80% of metabolites are excreted in the urine within 24 hours. However, the many variables which affect the absorption, metabolism and clearance of xylene include exercise, alcohol intake, cigarette smoking, co-exposure to other solvents, gender, and gastrointestinal, hepatic and renal pathology. Xylene in high concentrations acts as a narcotic, inducing neuropsychological and neurophysiological dysfunction. Respiratory tract symptoms are also frequent. More chronic, occupational exposure has been associated with anemia, thrombocytopenia, leukopenia, chest pain with ECG abnormalities, dyspnea and cyanosis, in addition to CNS symptoms. Concomitant exposure to xylene and other solvents, including toluene, affected hematological parameters, liver size, liver enzymes, auditory memory, visual abstraction, and vibration threshold in the toes. Normal metabolic pathways were altered and significant increases in some serum bile acids may reflect early liver damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗

Assessment of the hepatotoxicity of acute and short-term exposure to inhaled p-xylene in F-344 rats.

Due to the ubiquitous presence of p-xylene in air and the existing uncertainty regarding its hepatotoxic potential, we examined the effect of acute and short-term exposure to inhaled p-xylene on the liver. Male F-344 rats were exposed to 0 or to 1600 ppm p-xylene, 6 h/d, for 1 or 3 d. Exposure to inhaled p-xylene caused no histopathological evidence of hepatic damage and had little or no effect on the serum levels of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, ornithine carbamyl transferase, alkaline phosphatase, and total bilirubin. Exposure to p-xylene for 1 or 3 d resulted in an increase in relative liver weight on d 1 post-exposure. The concentration of hepatic cytochrome P-450 was increased by both p-xylene exposure regimens on d 1 postexposure and had returned to control levels by d 3 following the single p-xylene exposure and by d 2 following the 3-d exposure. These observations provide consistent evidence that acute and short-term exposure to 1600 ppm p-xylene by inhalation did not produce overt hepatotoxicity but resulted in a significant increase in the concentration of hepatic cytochrome P-450, the principal enzyme system involved in the metabolic biotransformation of xenobiotics.

Administration, Inhalation↗

Combined effects of xylene and alcohol on the central nervous system.

Ten healthy male students were exposed to m-xylene alone at concentrations of about 6 and 11.5 mumol/l and given a single dose of alcohol (0.4 and 0.8 g/kg) prior to exposure. The effects of these xylene concentrations and alcohol doses, as well as the combined effects of the two xylene concentrations with the higher alcohol dose on psychophysiological functions, such as body balance and reaction time, were assessed. Xylene alone did not significantly impair these functions, although there was a tendency towards impairment by the exposure to the higher xylene concentration. The impairment caused by alcohol alone was dose-dependent and exceeded that caused by xylene alone. The deleterious effects of xylene combined with alcohol were usually additive, although antagonism of alcohol effects on body balance by the higher xylene concentration was observed. The effects were pharmacodynamic rather than pharmacokinetic in nature.

Adult↗

Physiological properties of a Pseudomonas strain which grows with p-xylene in a two-phase (organic-aqueous) medium.

Pseudomonas putida Idaho utilizes toluene, m-xylene, p-xylene, 1,2,4-trimethylbenzene, and 3-ethyltoluene as growth substrates when these hydrocarbons are provided in a two-phase system at 5 to 50% (vol/vol). Growth also occurs on Luria-Bertani medium in the presence of a wide range of organic solvents. The ability of the organism to grow in the presence of organic solvents is correlated with the logarithm of the octanol-water partition coefficient, with dimethyl-phthalate (log P(OCT) = 2.3) being the most polar solvent tolerated. During growth with p-xylene (20% [vol/vol]), there was an initial lag period accompanied by cell death, which was followed by a period of exponential growth. The stationary phase of growth was characterized by a dramatic decrease in cell viability, although cell dry weight and turbidity measurements slowly increased. Electron micrographs revealed that during growth in the presence of p-xylene, the outer cell membrane becomes convoluted and membrane fragments are shed into the culture medium. At the same time, the cytoplasmic membrane invaginates, forming vesicles, and becomes disorganized. Electron-dense intracellular inclusions were observed in cells grown with p-xylene (20% [vol/vol]) and p-xylene vapors, which are not present in cells grown with succinate. Attempts to demonstrate the presence of plasmid DNA in P. putida Idaho were negative. However, polarographic studies indicated that the organism utilizes the same pathway for the degradation of toluene, m-xylene, and p-xylene as that used by P. putida mt-2 which contains the TOL plasmid pWWO.(ABSTRACT TRUNCATED AT 250 WORDS)

Benzene Derivatives↗