Genetic diversity of Escherichia coli recovered from the oral cavity of beef cattle and their relatedness to faecal E. coli.

AIMS: To determine the genetic diversity of generic Escherichia coli recovered from the oral cavities of beef cattle and their relatedness to E. coli isolated from the faeces of cattle during pasture grazing and feedlot finishing. METHODS AND RESULTS: A total of 484 E. coli (248 oral and 236 faecal isolates) were obtained from eight beef cattle after 1 and 5 months of grazing on pasture and after 1 and 5 months in a feedlot. The random amplification of polymorphic DNA (RAPD) method was used to genetically characterize these isolates. The RAPD patterns showed that ca 60% of E. coli recovered from the oral cavities and faeces during pasture and feedlot shared a close genetic relatedness. A number of E. coli with unique RAPD types were also found either in the oral cavities or faeces. Most of the E. coli RAPD types recovered from the oral cavities were shared among animals, but there were also RAPD types which were unique to individual animals. The E. coli populations of the oral cavities were genetically diverse and changed over time. CONCLUSIONS: This study indicates that there are large numbers of E. coli carried in the oral cavities of beef cattle and those E. coli are closely related to strains found in the faeces. The oral cavities of cattle harbour a genetically diverse E. coli population. SIGNIFICANCE AND IMPACT OF THE STUDY: The oral cavity may be an important reservoir of enteric pathogens which may transfer to meat during carcass dressing. A better understanding of the molecular ecology of E. coli in cattle would assist the design of approaches to control pathogenic strains during beef production and processing.

Genotypic analysis of Escherichia coli recovered from product and equipment at a beef-packing plant.

AIMS: To identify sources of Escherichia coli on beef by characterizing strains of the organism on animals, equipment and product at beef-packing plant. METHODS AND RESULTS: Generic E. coli were recovered from hides, carcasses, beef trimmings, conveyers and ground beef during the summer of 2001 (750 isolates) and winter of 2002 (500 isolates). The isolates were characterized by Random Amplification of Polymorphic DNA (RAPD). The numbers of E. coli recovered from dressed carcasses were less than the numbers recovered from hides. The numbers recovered from chilled carcasses were too few for meaningful analysis of the strains present on them but the numbers recovered from trimmings and ground beef were larger. The RAPD patterns showed that the majority of isolates from hides, carcasses, beef trimmings, conveyers and ground beef were of similar RAPD types, but a few unique RAPD types were recovered from only one of those sources. The E. coli populations present on the hides of incoming animals and in the beef-processing environment were highly diverse. Randomly selected E. coli isolates from each of the five sources were further characterized by pulsed-field gel electrophoresis (PFGE). Most genotypes of E. coli defined by PFGE corresponded to the E. coli types defined by RAPD. CONCLUSIONS: The hides of the incoming animals appeared to be only one of the sources of the E. coli on trimmings and in ground beef, as additional sources were apparently present in equipment used for carcass breaking. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that hazardous microbiological contamination of meat may occur after the dressing of carcasses at commercial beef-packing plants, which suggests that attention should be given to the control of the contamination of meat during carcass breaking as well as during the dressing of carcasses.

Efficacy enhancement of trisodium phosphate against spoilage and pathogenic bacteria in model biofilms and on adipose tissue.

A two-step approach for enhancing the efficacy of trisodium phosphate (TSP) was evaluated using meat spoilage and pathogenic bacteria in flow cell biofilms and adipose tissue model systems. The process was based on the plasmolysis of attached bacteria (biofilms) with a hyperosmotic solution (1.5 M NaCl) and the subsequent deplasmolysis of cells with a low-osmotic-strength solution containing different concentrations of TSP (0.1, 0.25, 0.5, 0.625, and 1.0 % [wt/vol]). Escherichia coli, Salmonella Enteritidis, Pseudomonas sp., Listeria monocytogenes, and Brochothrix thermosphacta strains were cultivated for 24 h as pure culture biofilms in glass flow cells with complex media and were then treated with either 0.1, 0.25, 0.5, 0.625, and 1.0% TSP, or the same TSP concentrations delivered in conjunction with plasmolysis-deplasmolysis (PDP). Confocal scanning laser microscopy, a commercial fluorescent viability probe, and image analysis were then used to quantify the relative abundances of living and dead cells remaining after the different treatment regimes. With the exception of L. monocytogenes (which was resistant to TSP concentrations of up to 5%), the PDP process increased the sensitivity of the test strains to TSP. However, when similar experiments were conducted with pork adipose tissue, it became evident that higher TSP concentrations were necessary to produce significant decreases in the number of viable cells and that the PDP process generally failed to enhance TSP efficacy. An exception was L. monocytogenes, which exhibited an increase in sensitivity to TSP when inoculated tissue was pretreated with 1.5 M NaCl. It is thought that factors contributing to the failure of the PDP process to enhance the activity of TSP in meat systems involves the mode of TSP antimicrobial activity, alkaline pH stress, and the chemically complex, buffered nature of meats. It remains to be determined whether the PDP process is suitable for use with other food grade antimicrobial agents or can be used in nonfood biofilm control applications.

Description of a simple detection assay for in situ production of bacteriocin on meat.

Using a modification of the agar diffusion assay, in situ bacteriocin production on meat was analyzed using cooked meat medium (CMM) and sterile pork tissue (lean and fat) with Carnobacterium piscicola LV17 as the producer and Carnobacterium divergens LV13 as the indicator strains. Contrary to what is observed in APT broth, bacteriocin production by C. piscicola LV17 occurred with growth at low inoculum levels (< or =10(4) CFU/cm2 or g of meat) on disks (10 cm2) of pork fat tissue (pH 6.58) and on CMM particles (pH 7.0) but not on disks of lean tissue (pH 5.61). The assays described in this study do not required sophisticated equipment and would be useful to study bacteriocin production on meat products stored under various conditions.

Integrating microbial decontamination with organic acids in HACCP programmes for muscle foods: prospects and controversies.

A considerable literature reports the antibacterial efficacy of dilute solutions of organic acids (lactic, acetic). With carcasses an overall reduction in surface contaminants of 1.5 log cycles can be expected. Carcass decontamination may not improve the safety of the resultant meat, but laboratory trials confirm that acid decontamination of subprimal and retail cuts is more efficacious. An advantage over many other intervention strategies is that residual antimicrobial activity is demonstrable over extended periods of storage. These studies have also shown that some meatborne pathogens are particularly sensitive to organic acids (i.e., Yersinia enterocolitica) while others are resistant (i.e., E. coli O157:H7). Dilute solutions of organic acids (1 to 3%) are generally without effect on the desirable sensory properties of meat when used as a carcass decontaminant. However, dependent on treatment conditions, lactic and acetic acid can produce adverse sensory changes when applied directly to meat cuts, with irreversible changes in appearance being a frequent occurrence. It is speculated that organic acid decontamination will be implemented in American abattoirs in an effort to meet specified performance standards for pathogen reduction as part of an overall HACCP program. In contrast, the EU advocates that strictly controlled processing hygiene is sufficient to ensure the safety of the product. Additional research is necessary to establish a set of treatment conditions that may permit a practicable reduction in bacterial contamination throughout the processing chain with a measurable effect on safety and storage life, without imposing any change in sensory properties. It will also be necessary to develop standard, objective measures to assess HACCP and the efficacy of decontamination procedures. Without such commercial studies controversy on the practicality of acid decontamination will persist.

The aerobic growth of Aeromonas hydrophila and Listeria monocytogenes in broths and on pork.

Flasks of tryptic soy broth (TSB), unacidified (pH 7.2) or acidified with HCl or lactic acid to pH 6.3 or 5.5, and samples of sterile pork fat or muscle tissue, were inoculated with logarithmic phase cultures of a strain of Aeromonas hydrophila or a strain of Listeria monocytogenes. The broth cultures were incubated at temperatures between 0 and 25 degrees C, and growth rates were determined from optical density increases. The tissue samples were incubated at temperatures between -2.4 and 25.2 degrees C, and growth rates were determined from viable count increases. Both organisms grew without lag in all broths at temperatures greater than 10 degrees C. A hydrophila did not grow at 5 degrees C in TSB acidified with lactic acid to pH 5.5, and grew in other broths at that temperature after a lag of about 10 h. The organism did not grow in either broth of pH 5.5 at 2 degrees C, but grew in other broths at that temperature after a lag of about 40 h. A hydrophila did not grow in any broth at 0 degrees C. L. monocytogenes grew in all broths at 5 degrees C only after a lag of about 60 h, and did not grow in any broth at 2 degrees C. For both organisms, the rates of growth, at any temperature, were lower in broths of pH 6.3, and lower again in broths of pH 5.5, than in the unacidified broth. Growth rates in the broths of pH 6.3 were similar, but growth rates were lower in lactic acid acidified broth of pH 5.5 than in HCl acidified broth of that pH. The data for the growth of each organism in each medium were well described by the regression line of the plot of the square roots of growth rates against temperature. A hydrophila grew on fat tissue of pH 6.3 +/- 0.3, without lag, at 1.8 degrees C and higher temperatures at rates greater than the rates of growth in unacidified TSB. Numbers of the organism declined on muscle tissue of pH 5.6 +/0 0.2 at any temperature. L. monocytogenes grew on fat tissue without lag at -0.3 degrees C and higher temperatures, at rates which at lower temperatures were greater than the rates of growth in unacidified TSB. The organism grew on muscle tissue only at temperatures greater than or equal to 15.4 degrees C, at rates which were less than the rates of growth in lactic acid acidified broth of pH 5.5. Models derived from the cultivation of A. hydrophila and L. monocytogenes in commercial broths appear to be highly unreliable guides to the behaviours of those organisms on pork.

Quality and bacteriological consequences of beef carcass spray-chilling: Effects of spray duration and boxed beef storage temperature.

The effects of water spray-chilling on beef carcass traits and muscle quality, bacteriology and retail case life were determined in a research abattoir. Chilling treatments were compared using 10 crossbred steer carcasses (280 ± 4 kg) at each spray duration (4, 8, 12 and 16 h) and each vacuum storage temperature (1, 4, 8 and 12 °C). Control sides were air-chilled (1 °C, 24 h) while spray-chilled sides were sprayed with an intermittent water mist at 1 °C in four, 60 s cycles/h for the initial 4-16 h of chilling. The effects of storage temperature were evaluated using vacuum packaged longissimus thoracis (LT) muscle at post-chill intervals of 2, 16, 30 and 44 days. Chilling treatment effects were similar at all spray-chill durations and LT vacuum storage times and temperatures. Carcass spray-chilling did not effect pH, lean colour, % moisture, sarcomere length, shear value or weight loss during the vacuum storage of LT muscle. Carcass fat colour tended to brighten as spray duration was extended up to 12 h, but there was a grey discoloration of fat at spray durations beyond 12 h. Chilling treatment had only marginal effects on anaerobic bacteria during the vacuum storage of LT muscles, or aerobic bacteria during the retail display of rib-eye steaks, and the retail case life of steaks was largely unaffected by spray-chilling. A linear relationship between spray-chill duration and carcass weight loss was determined and carcass shrinkage was reduced by 0.08 g/100 g for every hour of spray-chilling. It was estimated that a major beef processing abattoir could utilize spray-chilling to save more than 2000 kg daily in carcass shrinkage, without compromising quality or increasing spoilage losses.

Control of beef spoilage by a sulfide-producing Lactobacillus sake strain with bacteriocinogenic Leuconostoc gelidum UAL187 during anaerobic storage at 2 degrees C.

Chill-stored, vacuum-packaged beef inoculated with sulfide-producing Lactobacillus sake 1218 developed a distinct sulfide odor within 3 weeks of storage at 2 degrees C, at which time the bacteria had reached maximum numbers of 10(6) CFU cm(-2). Coinoculation of the meat with the wild-type, bacteriocinogenic (Bac+) strain of Leuconostoc gelidum UAL187 delayed the spoilage by L. sake 1218 for up to 8 weeks of storage. Coinoculation of meat samples with an isogenic, slowly growing Bac+ variant, UAL187-22, or with the Bac- variant UAL187-13 did not delay the onset of spoilage by L. sake 1218. The study showed that spoilage of chill-stored, vacuum-packaged beef by a susceptible target organism could be dramatically delayed by the Bac+ wild-type strain of L. gelidum UAL187. Inoculation with L. sake 1218 can be used as a model system to determine the efficacy of biopreservation of vacuum-packaged meats.

Effect of growth of selected lactic acid bacteria on storage life of beef stored under vacuum and in air.

The effect of growth of different types of lactic acid bacteria (LAB) on the storage life of normal pH beef was determined anaerobically (under vacuum) and aerobically. Four LAB from meat were inoculated separately onto sterile slices of lean beef. Inoculated samples were stored anaerobically at 2 degrees C for 10 weeks or stored aerobically in an oxygen permeable film at 7 degrees C for 10 days, with and without previous storage under vacuum at 2 degrees C. The LAB strains used were Carnobacterium maltaromicus (previously C. piscicola) LV17 and UAL26, Leuconostoc gelidum UAL187-22 and Lactobacillus sake Lb706. Storage life was determined by sensory panel evaluation of colour and odour. Under anaerobic conditions Lb. sake Lb706, inoculated at log 2 CFU/cm2, grew rapidly to reach maximum population within three weeks of storage. L. gelidum UAL187-22 also grew on the meat but at a slower rate. In contrast, growth of C. maltaromicus LV17 and UAL26 was unpredictable, achieving maximum population after 2 to 8 weeks. None of the test strains caused spoilage of the meat within the 10-week storage period under vacuum. When the test organisms were inoculated at an initial level of log 4 CFU/cm2, C. maltaromicus LV17 and UAL26 produced off-odours after 8 weeks of storage under vacuum at 2 degrees C. Under aerobic conditions at 7 degrees C, all four of the strains grew well on the beef samples. C. maltaromicus LV17 and UAL26 and Lb. sake Lb706 all caused off-odours and discoloration. The rate of aerobic deterioration in meat quality was faster with increased time of storage under vacuum. L. gelidum UAL187-22 could be a suitable antagonistic strain with the potential to extend the storage life of beef, stored anaerobically and packaged aerobically for retail sale, without producing undesirable sensory changes.

Lactic acid inhibition of the growth of spoilage bacteria and cold tolerant pathogens on pork.

The antibacterial effects of a 3% solution of lactic acid at 55 degrees C were assessed, by examining aerobic bacterial growth on artificially-inoculated pork fat and lean tissue. Discs of fat or lean tissues, each of 10 cm2 surface area, were aseptically excised from pork Longissimus dorsi muscle and inoculated with the cold tolerant pathogens Listeria monocytogenes 4b Scott A no. 3, Yersinia enterocolitica 0:4,32 or Aeromonas hydrophila ATCC 7966, or with the wild type spoilage bacteria Pseudomonas fragi or Brochothrix thermosphacta. After inoculation, each meat disc was immersed in water or lactic acid for 15 s and aerobic bacterial growth followed during 15 days of storage at 4 degrees C. P. fragi and B. thermosphacta grew on both fat and lean, but the pathogens grew on fat tissue only and A. hydrophila did not survive on lean. Lactic acid reduced all test bacteria on fat to below detectable levels within 4 days of treatment and no bacteria could be recovered from acid-treated fat surfaces for the remainder of the 15-day storage interval. Bacteria attached to lean were generally more resistant to lactic acid. In some instances the acid was bacteriostatic (P. fragi, L. monocytogenes) while in others the population declined at a greatly reduced rate as compared with a similar population on fat (B. thermosphacta, Y. enterocolitica). A. hydrophila was equally sensitive to lactic acid on lean and fat. Depending upon the tested strain, tissue type and storage time, maximum reductions in the number of bacteria recovered from acid treated pork ranged from 1 to 8 log cycles. The high bactericidal efficacy of lactic acid applied to pork fat was attributable to a low tissue pH, which varied from 3.49 to 4.41 during the 15 days of aerobic storage.

Bacteriology and Retail Case Life of Pork After Storage in Carbon Dioxide.

The effects of prolonged, anoxic storage, under CO2 at -1.5°C, upon the bacteriology and case life of pork on its subsequent transfer to the aerobic conditions of simulated retail display at 8°C was examined. Brochothrix thermosphacta , lactic acid bacteria, enterics, and pseudomonads were enumerated. Panel scores for odor and appearance acceptability were used to quantify retail case life. Lactic acid bacteria were the only bacteria found during loin storage in CO2 for up to 24 weeks. Those organisms reached maximum number of 107 CFU/cm2 within 9 weeks. The number of lactic acid bacteria initially found on the freshly cut surfaces of loin chops increased linearly during the first 9 weeks of loin storage in CO2. Thereafter, they continued to grow on the chops and dominated the spoilage flora during retail display. The pseudomonads grew rapidly and emerged as the next most numerous organism, while B. thermosphacta and enterics showed only limited aerobic growth. The acceptability of pork chop appearance and odor was adversely affected by loin storage time. Each 6-week interval of loin storage produced a 1 d reduction in case life. Should controlled atmospheres be a practicable means of meat distribution to the retail marketplace, efforts will be necessary to assure a maximum case life after their removal from preservative packagings.

Cryogenic chilling of pork carcasses: Effects on muscle quality, bacterial populations and palatability.

Two experiments were conducted to determine the effects of cryogenic chilling on the carcass shrinkage, meat quality, bacterial condition and palatibility of pork. In experiment I, pork sides were chilled at 1°C (n = 20), or immersed in liquid nitrogen (LN) for 1 or 3 min prior to placement in a 1°C cooler. Muscle temperature in the loin was significantly lower at 2 and 6 h post mortem in treated compared to control sides, and loin muscle pH was higher (P < 0·05) at 6 h post mortem in sides immersed for 3 min in LN. Carcass side shrinkage was reduced from 29·3 g kg(-1) in control sides to 20·9 and 13·5 g kg(-1) in sides dipped in LN for 1 and 3 min. Chilling treatment had no significant effect on the survival of mesophilic bacteria on carcass sides, on meat colour, drip loss, protein solubility or sarcomere length, but sides dipped for 1 min in LN has a higher muscle shear value than control sides. In experiment II, carcass sides from halothane positive (H+) and negative (H-) pigs were conventionally chilled (n = 49), immersed in LN for 3 min (n = 23), or electrically stimulated and chilled in LN for 3 min (n = 26). Similar results for temperature, pH, colour, protein solubility and drip loss in loin muscle were found to those in experiment I. Laboratory taste panel results showed that chilling treatment had no effect on palatability. Genotype produced meaningful differences in most palatability attributes with H+ pigs having less tender, less juicy and less desirable flavour than pork from H- pigs. Laboratory studies with inoculated fresh muscle slices showed that a 3 min immersion in LN resulted in a 10-fold reduction in the aerobic spoilage pseudomonads, but effects upon other spoilage bacteria and potential human pathogens were less pronounced. It was concluded that cryogenic chilling using LN reduced carcass shrinkage during cooling, but had no consistent effects on meat quality, palatability or bacterial numbers on the carcass. In contrast, genotype had a significant effect on most pork quality and palatability attributes.

Inability of a bacteriophage pool to control beef spoilage.

The biological control of beef spoilage, with a bacteriophage (phage) pool, was evaluated under simulated retail conditions. A pool of seven phages was selected with the potential to lyse 78% of 86 Pseudomonas test strains. Subsequent host range studies with 1023 pseudomonads from three meat species (beef, pork, lamb) and five abattoirs showed that 585 (57.2%) isolates were susceptible to the phage pool. Depending on bacterial origin, bacterial sensitivity to lysis by the phage pool varied from 25 to 72%. When added to ribeye steaks, the phage pool produced a significant reduction in Pseudomonas growth but this was not sufficient to produce any significant effect upon the retail shelf life of beef. The inability of phages to control beef spoilage was not attributed to a loss of phage virulence since sufficient densities (log pfu/cm2 = 5 to 6) of virulent phage could be re-isolated from beef, 14 days after treatment. It was concluded that the efficacy of the current phage pool was limited by a narrow range of specificity.

Morphology of Brochothrix thermosphacta phages.

Twenty-one Brochothrix thermosphacta phages were examined by electron microscopy. They had isometric heads and contractile or long and noncontractile tails, and were grouped into three species belonging to the Myoviridae (species A19) or Siphoviridae (species NF5 and BL3) families of tailed phages. Species A19 has highly characteristic morphological features and resembles well-known phages of Bacillus, Lactobacillus, Staphylococcus and Streptococcus, indicating phylogenetic relationships between these phages as well as their hosts.

Psychrotrophic Brocothrix thermosphacta bacteriophages isolated from beef.

A total of 15 wild-type Brocothrix thermosphacta strains isolated from beef and the type strain, B. thermosphacta ATCC 11509, were used as hosts for the isolation of bacteriophages under psychrotrophic conditions (7 degrees C). A total of 21 virulent, psychrotrophic phages were successfully isolated and purified from aqueous extracts of spoiled rib steaks. Phage plaque size and plating efficiency significantly increased as incubation temperature was reduced from 25 to 1 degree C. Electron microscopy of two homologous B. thermosphacta phages showed the virions to consist of hexagonal heads and tails, with terminal appendages clearly visible on one of the phages. On the basis of culture and biochemical data, the wild-type B. thermosphacta strains had characteristics identical to those of strain ATCC 11509. However, specific differences in the pattern of susceptibilities to the phages revealed the presence of 14 distinct phage lysotypes. Phage typing may provide a rapid and sensitive means of differentiating B. thermosphacta strains.

Inverse relationship between the susceptibility of lipopolysaccharide (lipid A)-pretreated mice to the hypothermic and lethal effect of lipopolysaccharide.

Mice pretreated (day 0) by a single injection of lipopolysaccharide (LPS) responded with hypothermic tolerance to (LPS) challenge on day 1 and with hypothermic hyperreactivity to LPS challenge on day 4. Reciprocally, mice pretreated similarly but with a higher challenge dose were hyperreactive with respect to LPS lethality on day 1, but highly tolerant to lethality when challenged on day 4. Hyperreactivity to LPS lethality (day 1) was evident from an accelerated onset of death as well as from a reduced 50% lethal dose in pretreated mice, the level of hyperreactivity being more pronounced with higher LPS pretreatment doses. Lethal hyperreactivity, however, was only seen after challenge with a 50% lethal dose of soluble LPS. In contrast, protection to lethality occurred after challenge with a 50% lethal dose of insoluble LPS (day 1). Tolerance to LPS lethality in mice was observed on day 4 after pretreatment with one (day 0) or four daily injections of LPS. Since reciprocal hyperreactivity (day 1) and cross-tolerance to lethality (day 4) could be achieved by treatment with Salmonella smooth- or rough-form LPS as well as with free lipid A, it was concluded that lipid A represents the active principle of LPS in inducing both hyperreactivity and tolerance to the lethal effect of LPS.

Lipid A-induced tolerance and hyperreactivity to hypothermia in mice.

Mice responded to lipopolysaccharide (LPS) with a dose-dependent, monophasic hypothermia reaching a maximum at 2 h postinjection. Degraded polysaccharide was not active; free lipid A, however, induced a similar pattern of hypothermia, indicating that the hypothermic principle of LPS was embedded within the lipid A component. The hypothermic response of mice to LPS was modified by prior exposure of the host to LPS. This altered reactivity was manifested by refractory periods (early and late tolerance), in which animals no longer responded with hypothermia, or a hyperreactive phase (hypersensitivity), in which hypothermic responses were greatly augmented upon LPS challenge. Thus, tolerance observed 24 h after a single injection of LPS (early tolerance) was followed, on further LPS challenge, by an enhanced hypothermic responses reaching a maximum on day 4. Further daily exposure of the animals to LPS eliminated hyperreactivity and led to the establishment of a late tolerance maximally expressed on day 8. Hyperreactivity could also be evoked on day 4 after a single injection of LPS. Mice pretreated with Salmonella S- and R-form LPS or free lipid A (Salmonella) demonstrated tolerance and hyperreactivity to both homologous and heterologous challenge. In addition, complete cross-tolerance was observed with S-form LPS derived from Shigella. It was concluded that the differential effects of LPS on host responses (tolerance and hyperreactivity) were due to lipid A.

Pseudomonas aeruginosa lipopolysaccharide: factors influencing toxicity for isolated mitochondria and endotoxin properties.

In the present study Pseudomonas aeruginosa lipopolysaccharide (LPS) exhibited the following endotoxin properties: (1) toxicity for mice; (2) gelation of the Limulus lysate; (3) induction of a localized Shwartzman reaction in the skin of rabbits, and (4) anticomplementary activity. Differences in LPS toxicity as measured with the rat liver mitochondrial assay system were found to be related to the nature of the bacterial growth media, the functional integrity of mitochondria, and the time and temperature of mitochondrial assay. The significance of these findings to P. aeruginosa infections is discussed, and it is concluded that LPS is a factor of importance.

Pseudomonas aeruginosa lipopolysaccharide: an uncoupler of mitochondrial oxidative phosphorylation.

The addition of Pseudomonas aeruginosa KCIIR LPS to respiring mitochondria stimulated the rate of substrate oxidation, reduced the respiratory control ratio, stimulated oxygen uptake in state 4, and released the inhibition imposed upon state 3 by atractyloside. It was concluded that LPS acted as an uncoupler of oxidative phosphorylation and that it produced effects similar to those observed with the classical uncoupler 2,4-dinitrophenol.