Rhodococci

Med Clin (Barc), 1991 Apr 13, 96(14), 541 - 3
{Rhodococcus equi pneumonia in a patient with acquired immunodeficiency syndrome}; de Quiros JF et al.; A male patient of 33 years of age with AIDS developed pneumonia with abscess formation and a slow clinical course . The sputum, bronchoalveolar lavage and blood cultures yielded a Gram-positive, partially acid-alcohol resistant bacillus which was identified as Rhodococcus equi . This organism has been uncommonly implicated in pneumonia in patients with immunodeficiency, particularly with neoplasia and severe cell immunity deficiency . It has been very rarely reported in patients with AIDS . To our knowledge, this is the first reported case in Spain.

J Bacteriol, 1991 Apr, 173(8), 2465 - 72
Cloning and characterization of genes responsible for metabolism of nitrile compounds from Pseudomonas chlororaphis B23; Nishiyama M et al.; The nitrile hydratase (NHase) of Pseudomonas chlororaphis B23, which is composed of two subunits, alpha and beta, catalyzes the hydration of nitrile compounds to the corresponding amides . The NHase gene of strain B23 was cloned into Escherichia coli by the DNA-probing method with the NHase gene of Rhodococcus sp . strain N-774 as the hybridization probe . Nucleotide sequencing revealed that an amidase showing significant similarity to the amidase of Rhodococcus sp . strain N-774 was also coded by the region just upstream of the subunit alpha-coding sequence . In addition to these three proteins, two open reading frames, P47K and OrfE, were found just downstream of the coding region of subunit beta . The direction and close locations to each other of these open reading frames encoding five proteins (amidase, subunits alpha and beta, P47K, and OrfE, in that order) suggested that these genes were cotranscribed by a single mRNA . Plasmid pPCN4, in which a 6.2-kb sequence covering the region coding for these proteins is placed under control of the lac promoter, directed overproduction of enzymatically active NHase and amidase in response to addition of isopropyl-beta-D-thiogalactopyranoside . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the cell extract showed that the amount of subunits alpha and beta of NHase was about 10% of the total cellular proteins and that an additional 38-kDa protein probably encoded by the region upstream of the amidase gene was also produced in a large amount . The 38-kDa protein, as well as P47K and OrfE, appeared to be important for efficient expression of NHase activity in E . coli cells, because plasmids containing the NHase and amidase genes but lacking the region coding for the 38-kDa protein or the region coding for P47K and OrfE failed to express efficient NHase activity.

Surg Neurol, 1991 Apr, 35(4), 321 - 4
A case of Rhodococcus equi brain abscess; Obana WG et al.; We treated a patient with acquired immunodeficiency syndrome for a brain abscess caused by Rhodococcus equi, an actinomycete that usually infects the lung in immunosuppressed hosts . Rhodococcus equi brain abscess is an extremely rare lesion that has never been reported in a patient with acquired immunodeficiency syndrome . The infection was cured by lengthy therapy with multiple antibiotics after aspiration of the lesion to identify the infective organism and determine its sensitivity to antibiotics.

Eur J Biochem, 1991 Mar 28, 196(3), 581 - 9
Characterization of a new cobalt-containing nitrile hydratase purified from urea-induced cells of Rhodococcus rhodochrous J1; Nagasawa T et al.; A new cobalt-containing nitrile hydratase was purified from extracts of urea-induced cells from Rhodococcus rhodochrous J1 in seven steps . At the last step, the enzyme was crystallized by adding ammonium sulfate . Nitrile hydratase was a 500-530-kDa protein composed of two different subunits (alpha subunit 26 kDa, beta subunit 29 kDa) . The enzyme contained approximately 11-12 mol cobalt/mol enzyme . A concentrated solution of highly purified nitrile hydratase exhibited a broad absorption spectrum in the visible range, with an absorption maxima at 410 nm . The enzyme had a wide substrate specificity . Aliphatic saturated or unsaturated nitriles as well as aromatic nitriles, were substrates for the enzyme . The optimum pH of the hydratase was pH 6.5-6.8 . The enzyme was more stable than ferric nitrile hydratases . The amino-terminal sequence of each subunit of R . rhodochrous J1 enzyme was determined and compared with that of ferric nitrile hydratases . Prominent similarities were observed with the beta subunit . However, the amino acid sequence of the alpha subunit from R . rhodochrous J1 was quite different from that of the ferric enzymes.

FEMS Microbiol Lett, 1991 Mar 1, 62(2-3), 277 - 80
Occurrence of two structural types of mercury reductases among gram-positive bacteria; Bogdanova ES et al.; Structural variants of mercury reductase containing the N-terminal domain, which is easily cleaved by trypsin, have been found in Gram-positive bacteria with a low genomic G + C content (Bacillus, Staphylococcus and, possibly, some other genera) . Mercury reductases without the N-terminal domain and relatively resistant to limited proteolysis are typical for Gram-positive bacteria with a high genomic G + C content (Arthrobacter, Citreobacterium, Micrococcus, Mycobacterium, Rhodococcus) . Both types of mercury reductase genes may be located on plasmids.

J Clin Microbiol, 1991 Mar, 29(3), 439 - 43
Identification of 15- to 17-kilodalton antigens associated with virulent Rhodococcus equi; Takai S et al.; Antigens of Rhodococcus equi were analyzed by immunoblotting with naturally infected foal sera . Immunoblots of whole-cell antigen preparations of clinical isolates of R . equi revealed that major protein bands with molecular masses of 15 to 17 kDa were present in all clinical isolates tested and all isolates virulent for mice . In contrast, the 15- to 17-kDa antigens were not identified by immunoblotting in ATCC 6939, a type strain of R . equi that was avirulent for mice . Whole-cell antigens of 102 environmental isolates were investigated by immunoblotting and the mouse pathogenicity test . Twenty-five of these isolates were demonstrated to contain the 15- to 17-kDa antigens by immunoblotting and were virulent for mice . The remaining 77 environmental isolates lacked the 15- to 17-kDa antigens and were avirulent for mice . These data suggest that the diffuse 15- to 17-kDa proteins are virulence-associated antigens with immunogenicity in foals and that they may be useful in marking virulent R . equi contamination in the environment of a horse-breeding farm.

Mikrobiologiia, 1991 Mar-Apr, 60(2), 334 - 8
{Ultrastructural features of Rhodococcus rubropertinctus and Streptococcus lactis dissociants}; Martynkina LP et al.; Three Rhodococcus rubropertinctus dissociants (R, S and M) and two Streptococcus lactis dissociants (R and S) were compared using the electron microscopy method of negative contrasting . The cell wall of R.rubropertinctus dissociants had a thickness of 40 nm (R), 30 nm (S) and 20 nm (M) . The cell wall of S . lactis dissociants was 35-55 nm (R) and 25-30 nm (S) thick . 1.5-2-fold variations in the thickness of cell walls could account for the different resistance of the dissociants against the action of such external factors as dehydration, antibiotics, UV, elevated NaCl concentrations, etc.

Vet Microbiol, 1991 Feb 15, 26(4), 323 - 33
Role of antibody to extracellular proteins of Rhodococcus equi in protection against R . equi pneumonia in foals; Machang'u RS et al.; Rhodococcus equi produces two exoenzymes (REE), a cholesterol oxidase in large amounts and a phospholipase C, which cause lysis of sheep red blood cells (SRBC) sensitized with Staphylococcus aureus beta toxin . Two immunization studies were done in foals to determine the role of antibody to REE in protection against R . equi pneumonia . In the first study, three foals (mean age 10 days) were vaccinated four times at 2-week intervals with over 1 million units of partially purified exoenzymes (PREE) . In the second study, three foals (mean age 19 days) were administered plasma from an adult horse vaccinated with PREE . Relatively low titres (16-32) of neutralizing antibody were detected in the foals of the former group, and passive transfer of neutralizing antibody (titres 32-64) occurred in the latter . Following immunization, principal foals and an equal number of similarly aged nonimmunized foals were challenged by aerosol with 1 x 10(10) live R . equi per day for 5 consecutive days . No severe clinical pneumonia developed in either group and, with one exception, only minor and resolving lung abscesses developed in these foals . These studies showed that antibody response of foals to immunization with PREE was poor, antibody to PREE did not prevent foals from developing lung abscesses following experimental infection, and that foals even as young as 3 weeks of age may be largely refractory to aerosol challenge with virulent R . equi.

J Am Acad Dermatol, 1991 Feb, 24(2 Pt 2), 328 - 32
Rhodococcus infection of the skin with lymphadenitis in a nonimmunocompromised girl; Martin T et al.; A 7-year-old girl had a 4 X 3 X 3 cm nodule on the left wrist with axillary lymphadenopathy . Acid-fast bacilli were seen on a smear from a biopsy specimen of this granulomatous skin lesion . A Rhodococcus species grew on culture . Skin infections caused by Rhodococcus may be more common than the few prior case reports suggest.

Scand J Infect Dis, 1991, 23(1), 1 - 6
Rhodococcus equi--an easily missed opportunistic pathogen; Doig C et al.; Over the last 10 years Rhodococcus equi has been identified as an important, albeit very rare, opportunistic pathogen . As the number of patients immunocompromised from HIV infection grows, this microorganism will likely become of increasing clinical importance . Infection with R . equi is usually insidious, causing progressive pulmonary disease that is typically pleural based at the time of microbiological diagnosis . As in the case we present, the clinical and microbiological diagnoses may be significantly delayed, either by the common pitfalls encountered in the laboratory identification of R . equi, or by the failure to recognize the pathogenic potential of the isolate . R . equi infection should be suspected in immunocompromised patients with pneumonia, when a pure or predominant growth of aerobic, non-sporeforming gram-positive bacilli is found on cultures of bronchoalveolar lavage fluid and other pulmonary pathogens have been excluded.

Rev Infect Dis, 1991 Jan-Feb, 13(1), 91 - 6
Rhodococcus equi infection in the patient with AIDS: literature review and report of an unusual case; Emmons W et al.; Rhodococcus equi is an aerobic, intracellular, gram-positive rod-coccus that is partially acid fast . The organism is primarily a pathogen in animals and has only rarely been seen in immunocompromised humans . Its most common manifestation is a slowly progressive pneumonia that may cavitate . Infections are thought to be acquired via respiratory exposure to animals or soil . R . equi infections are difficult to treat, usually requiring prolonged administration of parenteral antibiotics and often necessitating surgical drainage . A case of cavitary pneumonia and recurrent bacteremia with R . equi in a patient with AIDS is reported, and the current literature on R . equi infections in humans is reviewed.

Rev Infect Dis, 1991 Jan-Feb, 13(1), 139 - 45
Rhodococcus equi infection in patients with and without human immunodeficiency virus infection; Harvey RL et al.; Rhodococcus equi is an uncommon pathogen in humans that has occasionally been reported to cause infection in individuals with impaired cellular immunity . We summarize 30 previously published reports of human infection with R . equi and describe one additional case in a patient with AIDS . Eleven (35%) of the patients discussed in this report had AIDS or human immunodeficiency virus (HIV) infection, which is emerging as the leading cause of immunosuppression in cases of R . equi infection . Seventy-seven percent of all patients had pneumonia due to R . equi, and the infiltrate frequently cavitated . When HIV-infected patients were compared with those not infected with the virus, symptoms, age, and frequency of pneumonia were similar . Sputum and blood cultures were more likely to be positive in HIV-infected patients . Individuals with HIV infection also had a higher incidence of simultaneous secondary infections and higher mortality than non-HIV-infected patients (54.5% vs . 20%) . The rate of survival for all patients was 75% when antibiotics were combined with surgical resection of infected tissue; in comparison, the survival rate among patients receiving antibiotics alone was 61.1%.

Clin Microbiol Rev, 1991 Jan, 4(1), 20 - 34
Rhodococcus equi: an animal and human pathogen; Prescott JF; Recent isolations of Rhodococcus equi from cavitatory pulmonary disease in patients with AIDS have aroused interest among medical microbiologists in this unusual organism . Earlier isolations from humans had also been in immunosuppressed patients following hemolymphatic tumors or renal transplantation . This organism has been recognized for many years as a cause of a serious pyogranulomatous pneumonia of young foals and is occasionally isolated from granulomatous lesions in several other species, in some cases following immunosuppression . The last decade has seen many advances in understanding of the epidemiology, pathogenesis, diagnosis, treatment, and immunity to infection in foals . The particular susceptibility of the foal is not understood but can be explained in part by a combination of heavy challenge through the respiratory route coinciding with declining maternally derived antibody in the absence of fully competent foal cellular immune mechanisms . R . equi is largely a soil organism but is widespread in the feces of herbivores . Its growth in soil is considerably improved by simple nutrients it obtains from herbivore manure . About one-third of human patients who have developed R . equi infections had contact in some way with herbivores or their manure . Others may have acquired infection from contact with soil or wild bird manure . R . equi is an intracellular parasite, which explains the typical pyogranulomatous nature of R . equi infections, the predisposition to infection in human patients with defective cell-mediated immune mechanisms, and the efficacy of antimicrobial drugs that penetrate phagocytic cells.

Zentralbl Mikrobiol, 1991, 146(6), 425 - 34
{Biosensors for the determination of phenol and benzoate on the basis of Rhodococcus cells and enzyme extracts}; Riedel K et al.; An amperometric biosensor for determination of phenol, cresol, benzoate and 2-methyl-4-chlorophenol using Rhodococcus-cells and enzyme extracts of Rhodococcus has been developed . The influence of cultivation of Rhodococcus-cells and preincubation of the biosensor with desired substrate on sensibility and specificity was been investigated . In relation to cultivation and preincubation the Rhodococcus sensor was high specific to benzoate or phenol and cresol . A linear range was obtained for phenol and benzoate up to 80 mumol and for 2-methyl-4-chlorophenol up to 400 mumol . The biosensor using enzyme extracts show a higher specificity, it is but necessary NADPH . A further disadvantage is the little measuring range of this sensor.

Microbiol Immunol, 1991, 35(3), 175 - 85
Mycolic acid-containing glycolipid as a possible virulence factor of Rhodococcus equi for mice; Gotoh K et al.; By the use of various Rhodococcus equi strains differing in the length of carbon chains of glycolipid, we examined whether the glycolipid, glucose monomycolate, was contributing to the virulence of R . equi for mice . R . equi strains with longer carbon chain mycolic acid showed a higher virulence as determined by lethality and granuloma formation in mice than those with shorter ones . When purified glycolipid was injected into mice, granuloma formation and liver damage were most prominent with the glycolipid having longer carbon chain mycolic acid . Only a representative strain with longer carbon chain mycolic acid persisted in the spleen of mice after intravenous injection, while a strain with shorter carbon chain mycolic acid was readily eliminated . These results suggested that glycolipid was at least one of the virulence factors of R . equi and that the carbon chain length of mycolic acid might be critical in the expression of virulence.

Arch Microbiol, 1991, 157(1), 76 - 9
Catabolism of 1,3-dinitrobenzene by Rhodococcus sp . QT-1; Dickel O et al.; The 1,3-dinitrobenzene-degrading Rhodococcus strain QT-1 was isolated under nitrogen limiting conditions from contaminated soil samples . Experimental data indicate tha 1,3-dinitrobenzene is metabolized via 4-nitrocatechol . Both compounds were oxidized by resting cells and nitro groups were completely eliminated as nitrite . Strain QT-1 utilizes both 1,3-dinitrobenzene and 4-nitrocatechol as source of nitrogen in the absence as well as in the presence of high amounts of ammonia . Growth on 4-nitrocatechol does not induce the enzyme(s) for the initial oxidation of 1,3-dinitrobenzene.

J Reprod Fertil Suppl, 1991, 44, 571 - 8
Protection against naturally acquired Rhodococcus equi pneumonia in foals by administration of hyperimmune plasma; Madigan JE et al.; A 2-year field study was performed to determine the capability of increasing Rhodococcus equi specific antibody in foals via plasma transfusion or mare vaccination, to determine the kinetics of R . equi (ELISA) antibody decay and to assess the protective effects of these procedures in foals on a farm endemic for R . equi . Plasma donors were vaccinated with a killed R . equi bacterin and produced high levels of anti-R . equi antibodies, which were harvested by plasmapheresis . In Experiment 1, 68 foals were given 1 litre of hyperimmune plasma intravenously (i.v.) between 1-60 days of age . Foal plasma R . equi antibody was significantly increased and high levels of R . equi antibody (ELISA) were maintained for 60 days . No R . equi pneumonia developed in any foals receiving plasma . In Experiment 2, 99 pregnant mares were vaccinated with R . equi bacterin at 30, 60 and 90 days before foaling . Group 1 foals (101:85 from R . equi immunized mares) also received plasma transfusions and Group 2 foals (14), from R . equi immunized mares, did not receive plasma transfusions . Pregnant mare immunization increased colostrum R . equi antibody significantly . Eight foals showed failure of transfer of specific R . equi antibody . The incidence of R . equi pneumonia was 2.9% in Group 1 foals and 43% in Group 2 foals . Vaccination of pregnant mares did not provide protection against R . equi pneumonia; however, plasma transfusion with hyperimmune plasma administered prior to R . equi exposure was significantly protective in foals.

Biodegradation, 1991, 2(1), 25 - 31
Dechlorination of pentachlorophenol by membrane bound enzymes of Rhodococcus chlorophenolicus PCP-I; Uotila JS et al.; Dechlorination (para-hydroxylation) of pentachlorophenol (PCP) and tetrachloro-para-hydroquinone (TeCH) and O-methylation of TeCH were demonstrated in cell extracts of Rhodococcus chlorophenolicus PCP-I . PCP para-hydroxylating activity was membrane bound, whereas TeCH dechlorinating enzyme was soluble . The PCP para-hydroxylating enzyme was solubilized by Triton X-100 and the requirement for both FAD and NADPH was shown . The dechlorinating activities were inducible in contrast to the constitutive TeCH O-methylating activity . The PCP para-hydroxylation was inhibited by its product TeCH, by anoxic conditions, and by different inhibitors of P450 . Participation of this cytochrome in the PCP hydroxylation was confirmed by the appearance of a carbon monoxide dependent peak of absorbance at 457 nm in the membrane fraction prepared from PCP degrading cells.

FEBS Lett, 1990 Dec 17, 277(1-2), 112 - 4
Detection of covalent enzyme-substrate complexes of nitrilase by ion-spray mass spectroscopy; Stevenson DE et al.; Nitrilase from Rhodococcus ATCC 39484 was found to consist of two species of Mr 40,258 +/- 2 and 40,388 +/- 2 Da . When the enzyme was incubated with nitrile substrates and the reaction quenched with acid, higher Mr species were observed . The mass differences were consistent with addition of a substrate molecule to each species . These results represent the first reported demonstration that this, or any other nitrilase forms a covalent intermediate with its substrates . The observation that the intermediate, suggested to be either a thioimidate or an acylenzyme, can be trapped by acidification indicates that the rate of breakdown of the intermediate is rate-limiting.

Zentralbl Bakteriol, 1990 Dec, 274(3), 299 - 315
Classification and identification of rhodococci; Goodfellow M et al.; Ninety-nine representative strains of Rhodococcus were compared through 129 unit characters and the data examined using the simple matching, Jaccard and pattern coefficients . Clustering was achieved using the unweighted pair group method with arithmetic averages algorithm . The numerical classification was not affected by the statistics used or by test error, estimated at 1.95% . Sixteen multimembered and 23 single member clusters were defined by the simple matching coefficient at or above the 89% similarity level . The numerical phenetic data support the taxonomic integrity of all 13 validly described species of Rhodococcus, including Rhodococcus aichiensis . Rapid enzymic tests based upon fluorophores 7-amino-4-methylcoumarin and 4-methylumbelliferone provide valuable data for rhodococcal systematics.

Zentralbl Bakteriol, 1990 Nov, 274(2), 203 - 13
Characterization of Nocardia, Rhodococcus and Gordona species by in vitro susceptibility testing; Boiron P et al.; Representative strains of Gordona, Nocardia and Rhodococcus were tested against 26 antimicrobial agents using the disc diffusion method . A distinct susceptibility profile was noted for most species . Nocardia asteroides, N . brasiliensis, N . otitidiscaviarum, N . transvalensis and N . vaccinii were rarely susceptible to the antibiotics tested; N . brevicatena and N . farcinica varied in their susceptibility depending on the strain tested . Only the antibiotic amikacin was active against all nocardiae . In contrast, the Gordona and Rhodococcus strains showed considerable susceptibility, in particular to the beta-lactam antibiotics . Amoxicillin + clavulanic acid and gentamicin were active against all of the gordonae and rhodococci . The differences in susceptibility patterns may prove useful in characterizing the genera Gordona, Nocardia and Rhodococcus and in separating species within these taxa.

J Am Vet Med Assoc, 1990 Oct 15, 197(8), 1039 - 42
Septicemia, atrial fibrillation, cardiomegaly, left atrial mass, and Rhodococcus equi septic osteoarthritis in a foal; Collatos C et al.; A foal with vegetative bacterial endocarditis affecting the wall of the left atrium was treated successfully with cefotaxime, erythromycin, and rifampin . Bacterial isolates included Escherichia coli from blood and Rhodococcus equi from a P-type osteomyelitic lesion of the left third metatarsal bone and from synovial fluid from the left metatarsophalangeal joint . Cardiac complications included cardiomegaly and atrial fibrillation, which responded to treatment with digoxin and quinidine sulfate . Cardiac function was considered normal 18 months after treatment . Bacteriologic cure of osteoarthritis was achieved by use of surgical debridement, lavage, and local and systemic antimicrobial treatment; however, lameness developed 18 months after treatment when training for flat racing was begun . Radiography revealed chronic degenerative joint disease.

Carbohydr Res, 1990 Oct 10, 206(2), 311 - 32
Structural analysis of the specific capsular polysaccharide of Rhodococcus equi serotype 2; Severn WB et al.; The specific capsular polysaccharide produced by Rhodococcus equi serotype 2 is a high-molecular-weight acidic polymer composed of D-glucose, D-mannose, D-glucuronic acid and 3-O-{(S)-1-carboxyethyl}-L-rhamnose in equimolar proportions . Structural analysis, employing a combination of chemical and n.m.r . techniques, established that the polysaccharide is composed of linear repeating tetrasaccharide units . (formula; see text) in which the beta-D-mannose residues carry O-acetyl groups at O-2 and O-3 to the extent of 1.7 mol equivalents . Unequivocal determination of the absolute chirality of the 3-O-{(S)-1-carboxyethyl}-alpha-L-rhamnose residues was achieved by chemical correlation with an authentic synthetic sample . The 1H and 13C-n.m.r . resonances of the native and O-deacetylated serotype 2 polysaccharides were fully assigned by homo- and heteronuclear chemical-shift correlation methods.

J Antibiot (Tokyo), 1990 Oct, 43(10), 1316 - 20
Nitrilase-catalyzed production of pyrazinoic acid, an antimycobacterial agent, from cyanopyrazine by resting cells of Rhodococcus rhodochrous J1; Kobayashi M et al.; Using resting cells of Rhodococcus rhodochrous J1, in which a large amount of nitrilase is induced, a simple and efficient bioconversion process for the production of pyrazinoic acid, an antimycobacterial agent, through catalysis by a nitrilase was developed . The reaction conditions for production of pyrazinoic acid were optimized . Under optimum conditions, 3.5 M cyanopyrazine was converted to pyrazinoic acid, with a molar conversion yield of 100% . The highest yield achieved corresponded to 434 g of pyrazinoic acid per liter of reaction mixture . The synthesized pyrazinoic acid was isolated and identified physico-chemically.

Am J Vet Res, 1990 Oct, 51(10), 1608 - 15
Production and partial characterization of monoclonal antibodies to the neotype strain of Mycobacterium bovis; Kuchinka GD et al.; Six monoclonal antibodies (MAB) to virulent Mycobacterium bovis ATCC 19210 were produced, using a suspension of heat-inactivated whole cells . Immunoglobulin isotype for MAB VMB6, VMB73, and VMB93 was IgG1, and for VMB31, VMB99, and VMB119, it was IgG2a . Monoclonal antibodies were examined for cross-reactivity to M tuberculosis, M kansasii, M fortuitum, M paratuberculosis, M avium serovars 1, 2, 4, 8, and 10, M chelonei, M phlei, M scrofulaceum, M smegmatis, Nocardia asteroides, and Rhodococcus equi . Monoclonal antibodies could be grouped on the basis of binding activity by ELISA and immunoblot analysis, in which MAB VMB6, VMB31, and VMB119 had binding activity to M bovis; MAB VMB93 and VMB99 detected M bovis and M tuberculosis antigens, and MAB VMB73 reacted with other mycobacterial species, as well as with N asteroides and R equi . Apparent molecular mass of antigens was 30 to 25 kilodaltons (kD) for VMB6, VMB31, and VMB119 and 63 kD for VMB93 and VMB99, and ranged from greater than 200 to 31 kD for VMB73, as estimated by immunoblot analysis . Monoclonal antibody binding activity to 18 field isolates of M bovis was evaluated, using ELISA . Each of 18 field isolates was detected, using MAB VMB6, VMB31, or VMB119; 10 isolates were detected, using MAB VMB93/VMB99, and 14 were detected by use of MAB VMB73 . Use of MAB in ELISA failed to detect antigens from M bovis strain AN-5.

Appl Microbiol Biotechnol, 1990 Oct, 34(1), 42 - 6
Aryl acylamidase from Rhodococcus erythropolis NCIB 12273; Vaughan PA et al.; A Rhodococcus erythropolis strain was isolated from soil on the basis of its ability to use acetaminophen as the sole source of both carbon and energy for growth . When grown in a complex medium containing an anilide inducer compound, the bacterium exhibited aryl acylamidase (EC 3.5.1.13) activity . This activity was not subject to carbon or nitrogen repression by the growth medium constituents as the enzyme was present throughout the exponential growth phase . The anilide was converted to the corresponding aniline, which was not further degraded . The enzyme was partially purified by a variety of methods including a batch ion exchange procedure, column ion exchange chromatography and hydrophobic interaction chromatography . The enzyme had a maximum activity at around pH 8.0 and had a Km for acetaminophen of 0.11 mM . Electrochemical assays of aryl acylamidase activity are described . The enzyme is suitable for use as a reagent in the clinical diagnostic measurement of acetaminophen.

J Bacteriol, 1990 Sep, 172(9), 4807 - 15
Purification and characterization of a novel nitrilase of Rhodococcus rhodochrous K22 that acts on aliphatic nitriles; Kobayashi M et al.; A novel nitrilase that preferentially catalyzes the hydrolysis of aliphatic nitriles to the corresponding carboxylic acids and ammonia was found in the cells of a facultative crotononitrile-utilizing actinomycete isolated from soil . The strain was taxonomically studied and identified as Rhodococcus rhodochrous . The nitrilase was purified, with 9.08% overall recovery, through five steps from a cell extract of the stain . After the last step, the purified enzyme appeared to be homogeneous, as judged by polyacrylamide gel electrophoresis, analytical centrifugation, and double immunodiffusion in agarose . The relative molecular weight values for the native enzyme, estimated from the ultracentrifugal equilibrium and by high-performance liquid chromatography, were approximately 604,000 +/- 30,000 and 650,000, respectively, and the enzyme consisted of 15 to 16 subunits identical in molecular weight (41,000) . The enzyme acted on aliphatic olefinic nitriles such as crotononitrile and acrylonitrile as the most suitable substrates . The apparent Km values for crotononitrile and acrylonitrile were 18.9 and 1.14 mM, respectively . The nitrilase also catalyzed the direct hydrolysis of saturated aliphatic nitriles, such as valeronitrile, 4-chlorobutyronitrile, and glutaronitrile, to the corresponding acids without the formation of amide intermediates . Hence, the R . rhodochrous K22 nitrilase is a new type distinct from all other nitrilases that act on aromatic and related nitriles.

J Am Vet Med Assoc, 1990 Sep 1, 197(5), 608 - 12
Surgical management of Rhodococcus equi metaphysitis in a foal; Desjardins MR et al.; A chronic Rhodococcus equi metaphysitis involving the distal growth plate of the left third metatarsal bone had induced a longstanding lameness in a young foal . Abnormal hematologic values included mild anemia, hyperfibrinogemia, mild leukocytosis, and neutrophilia . Radiography of the distal portion of MT3 revealed a radiolucent zone on the medial aspect of the growth plate, and small pieces of bone suggestive of sequestra . Treatment with erythromycin estolate and rifampin, aggressive surgical debridement, and cancellous bone grafting helped resolve the bone infection.

J Clin Microbiol, 1990 Sep, 28(9), 2142 - 4
Value of the O-nitrophenyl-beta-D-galactopyranoside test to differentiate among the aerobic actinomycetes; Flores M et al.; A comparative study to determine beta-D-galactosidase activity among 171 strains of aerobic actinomycetes (including mycobacteria and rhodococci) was performed by using two growth media and four O-nitrophenyl-beta-D-galactopyranoside (ONPG) substrates . The ONPG test was found to be a valuable screening test to differentiate between the ONPG-positive Nocardia spp . and the rapidly growing ONPG-negative mycobacteria and rhodococci . However, ONPG results varied significantly depending on the growth medium and test substrate used.

Vet Microbiol, 1990 Aug, 24(2), 205 - 9
The development and evaluation of an enzyme-linked immunosorbent assay for the detection of Mycobacterium bovis; Duffield BJ; A double antibody sandwich layer enzyme-linked immunosorbent assay (ELISA) was used to detect Mycobacterium bovis . The ELISA detected M . bovis is pure culture at concentrations of 1 x 10(5) colony-forming units (CFU) ml-1 and greater, compared to a minimum detection level of 1 x 10(6) CFU ml-1 for isolation techniques . Neither technique detected M . bovis at 1 x 10(4) CFU ml-1 . The ELISA did not cross-react with common mycobacterial contaminants such as Mycobacterium avium intracellulare-scrofulaceum complex serotypes 18 and 42, M . terrae, M . fortuitum, M . flavescens and with Escherichia coli or Rhodococcus equi . Further work is needed to evaluate this assay in detecting M . bovis in tissues and the environment.

J Gen Microbiol, 1990 Jul, 136 ( Pt 7), 1357 - 63
Structure of a Rhodococcus gene encoding pigment production in Escherichia coli; Hart S et al.; A 2.1 kbp DNA fragment from Rhodococcus strain ATCC 21145 gave rise to the production of blue and pink pigments in Escherichia coli when cloned downstream of a strong promoter . The sequence of this DNA fragment contains a single open reading frame with a putative ribosome-binding site, potentially coding for a single protein of Mr 42,560 . Deletion analysis and in vitro transcription-translation experiments support the hypothesis that pigment production in E . coli is due to a single enzyme whose catalytic activity is still unknown . This small pigment gene may become useful for the development of a new generation of chromogenic cloning vectors which do not require expensive substrates for the detection of gene expression.

Cesk Epidemiol Mikrobiol Imunol, 1990 Jul, 39(4), 228 - 36
{Isolation of Listeria ivanovii in Slovakia}; Elischerova K et al.; From October 1977 to May 15, 1989 in Slovakia 39 strongly haemolytic strains of L . ivanovii were isolated from a woman after delivery of a stillborn foetus--from the lochiae, placenta and rectal smear, from five symptom-free subjects from the faeces, from the rectal smears of two sheep, from the intestinal contents in the portion of the terminal ileus from 28 free living small terrestrial mammals, one strain from meat--beef steak, from the lungs, liver and kidneys of a dead young sheep and one strain from silage . The strains from the lochiae and placenta as well as those from the organs of sheep were detected already as primocultures on commonly used blood agar, the other strains after varying periods of cold enrichment and subsequent inoculation on a solid selective medium with nalidixic acid and acriflavin . All strains had typical biochemical properties, among which from the diagnostic aspect in particular the catalase formation and acid formation from d-xylose is important, as well as the negative reaction with mannitol, rhamnose and alpha-methyl-d-mannoside . All strains agglutinated in O-factor serum X and gave a typical shovel-like shape CAMP test with Rhodococcus equi . The frequent incidence of L . ivanovii in micromammals indicates their importance as reservoir animals for this species of microorganism . The latter were detected on the territory of all three Slovak regions and in the capital of Slovakia, Bratislava.

Zentralbl Bakteriol, 1990 Jun, 273(2), 179 - 83
Functional similarity of Listeria ivanovii and Staphylococcus aureus in CAMP test; Brzin B et al.; On the basis of synergistic haemolysis of Listeria ivanovii and Rhodococcus equi, we suspected CAMP positiveness of R . equi and complete CAMP negativeness of L . ivanovii and confirmed that the latter, showing a double-zone haemolysis like Staphylococcus aureus, could produce the same phenomenon in the CAMP test if used instead of staphylococci . This functional similarity of L . ivanovii and CAMP Staphylococcus could be used as an additional diagnostic test for L . ivanovii.

Res Microbiol, 1990 Jun, 141(5), 599 - 607
{Chemotaxonomy of gram-positive bacteria metabolizing beta-caryophyllene}; Asselineau J et al.; Chemotaxonomic identification of coryneform bacteria metabolizing b-caryophyllene was attempted . The following phospholipids were identified as main components of the bacterial extracts: cardiolipids, phosphatidylethanolamine, phosphatidylinositol and mannosides of phosphatidylinositol . Saponification of the lipid extracts gave a mixture of hydroxylated and nonhydroxylated fatty acids . Among the latter, oleic and tuberculostearic acids were identified . The hydroxylated fatty acids were analysed by thin-layer chromatography and mass spectrometry (as methyl esters) . From the results thus obtained, the strains appeared to be more closely related to the genus Rhodococcus than to the genus Nocardia.

Am J Vet Res, 1990 May, 51(5), 766 - 71
Influence of Rhodococcus equi on the respiratory burst of resident alveolar macrophages from adult horses; Brumbaugh GW et al.; Opsonized Rhodococcus equi activated the respiratory burst of resident alveolar macrophages (AM) from adult horses in a logarithmic-linear, mass-related manner . The effect of R equi was not significantly different from that of equal masses of opsonized zymosan A . Therefore, R equi does not appear to attenuate the respiratory burst of equine AM . The stimulatory effect of R equi was not reflected by increased production of superoxide anion (O2-), but increased activity of the hexose monophosphate shunt was observed . These results suggest a similarity between the respiratory burst of AM from horses and that of AM from rabbits . We concluded that resident AM from adult horses do not produce O2- concurrently with an increase in activity of the hexose monophosphate shunt when stimulated with either opsonized zymosan A or opsonized R equi . This suggests that O2- is not an important component of the antibacterial defenses of equine AM . Whether equine AM are incapable of producing O2- or require different stimuli to produce it was not determined.

Plasmid, 1990 May, 23(3), 242 - 7
Nocardioform arsenic resistance plasmids and construction of Rhodococcus cloning vectors; Dabbs ER et al.; One of a number of large nocardioform plasmids previously obtained by a primarily genetic approach was reduced in size to about approximately 11 kb . This smaller plasmid possessed determinants for resistance to sodium arsenate and sodium arsenite, as well as immunity to nocardiophage Q4 . It was joined to an Escherichia coli-positive selection vector constructed by M . Zabeau and colleagues, which had the EcoR1 endonuclease gene placed under the control of the PR promoter of lambda as well as a bla determinant . The resulting shuttle vector of about 14.6 kb was maintained in E . coli and in several strains of Rhodococcus . The vector was efficient in cloning DNA without prior alkaline phosphatase treatment, as a result of the presence of the positive selection function . This function was not significantly expressed in Rhodococcus, and the presence of the nocardioform resistance determinants led to no increase in arsenate or arsenite resistance in E . coli . The presence of the bla gene resulted in an increase of about threefold in ampicillin resistance in Rhodococcus strains.

Mikrobiol Zh, 1990 May-Jun, 52(3), 63 - 5
{The mitogenic properties of Fusarium graminearum and Rhodococcus erythropolis enzymes}; Fedorovskaia EA et al.; Blast transformation of peripheral blood lymphocytes in healthy people is studied by enzymes of the microbic origin possessing the lectin activity . Galactose oxidase of Fusarium graminearum IMV-F-1060 is shown to be mitogenically active with respect to the lymphocytes in the culture in vitro and may be one of home sources of lymphocytic mitogens for the laboratory investigations.

Biochem Biophys Res Commun, 1990 Apr 30, 168(2), 437 - 42
Purification of inactivated photoresponsive nitrile hydratase; Nagamune T et al.; Photoresponsive nitrile hydratase from Rhodococcus sp . N-771 was purified in its inactivated form . The enzyme had a molecular weight of approximately 60 kDa and consisted of 2 subunits each having molecular weight of 27.5 and 28 kDa . The enzyme also contained 2 iron atoms/enzyme as a cofactor . The enzyme was more stable in its inactivated form, rather than the activated during storage in the dark . The enzyme was most stable in the temperature region of 0-35 degrees C, and lost its activity above 40 degrees C . The enzyme was most stable in the pH region of 6-8 . The optimum temperature and pH for the enzyme activity was 30 degrees C and 7.8, respectively . The enzyme showed wide substrate specificity, and most of the metal ions did not affect enzyme activity significantly . The absorption spectrum revealed the presence of some cofactor which changed form after photoirradiation.

Chest, 1990 Apr, 97(4), 1000 - 1
Pulmonary malacoplakia and Rhodococcus equi infection in a patient with AIDS; Scannell KA et al.; We describe a case of pulmonary malacoplakia and Rhodococcus equi cavitary pneumonia in a patient with AIDS . The relationship between Rhodococcus equi, a rare bacterial human pathogen, and malacoplakia, an unusual type of chronic granulomatous inflammation, is discussed.

Biochem Cell Biol, 1990 Apr, 68(4), 778 - 89
Structural analysis of the specific capsular polysaccharide of Rhodococcus equi serotype 1; Leitch RA et al.; The specific capsular polysaccharide produced by Rhodococcus equi serotype 1 was found to be a high molecular weight acidic polymer composed of D-glucose, D-mannose, and D-glucuronic acid . Structural analysis of the polysaccharide employed a combination of chemical and nuclear magnetic resonance techniques, from which it was determined that the polysaccharide possessed a linear repeating tetrasaccharide unit containing a single O-acetyl substituent and and acetal-linked pyruvic acid moiety: {formula: see text} The 1H and 13C nuclear magnetic resonances of O-deacetylated and pyruvic-free serotype 1 polysaccharides were fully assigned by homo- and hetero-nuclear chemical shift correlation methods.

J Clin Microbiol, 1990 Feb, 28(2), 328 - 31
Use of partially purified 54-kilodalton antigen for diagnosis of nocardiosis by Western blot (immunoblot) assay; Boiron P et al.; A Western blot (immunoblot) assay is presented for the diagnosis of nocardiosis with a specific immunodominant 54-kilodalton (kDa) antigen purified from a culture filtrate of Nocardia asteroides by immunoaffinity chromatography . The chromatography column was prepared with immunoglobulin G obtained from sera from patients with lepromatous leprosy . Unbound solutes consisted of specific, partially purified N . asteroides antigens, primarily a 54-kDa band, accompanied by two others of 31 and 62 kDa . The Western blot technique was applied to detecting the immunologic response to nocardiae . Immunodetection was performed according to the biotin-avidin system, which greatly improved the detection of antibodies even in immunosuppressed hosts . Each of 16 serum samples from immunosuppressed or immunocompetent patients infected with N . asteroides reacted with the 54-kDa band, and two reacted with the 31- and 62-kDa bands . Each of the serum specimens obtained from patients with mycetoma caused by Nocardia brasiliensis or Rhodococcus rhodochrous reacted with the 54-kDa band . There was no reaction to either the 54- or the 31-kDa antigen with all serum samples obtained from patients with tuberculosis, except one, with all serum samples obtained from patients with leprosy, or with all sera obtained from healthy controls . The 54-kDa protein is a candidate to be used as a probe to study the humoral immunologic response to nocardiae.

Antonie Van Leeuwenhoek, 1990 Jan, 57(1), 33 - 6
Kinetic studies of phenol degradation by Rhodococcus sp . P1 . II . Continuous cultivation; Hensel J et al.; The degradation of phenol by Rhodococcus sp . P1 was studied in continuous culture systems . The organism could be adapted by slowly increasing concentration, step by step, up to 30.0 g.l-1 phenol in the influent . The degradation rate reached values of about 0.3 g.g dry mass-1.h-1 . Large step increases in phenol concentration and addition of further substrates (e.g., catechol) were tolerated up to a certain concentration . With increasing dilution rate and increasing inlet phenol concentration the stability of the system decreased . Nomenclature: D--Dilution rate, h-1, Dc--Critical dilution rate, h-1, Dx--Yield, g dry mass.l-1.h-1, Ks--Monod saturation constant, g.l-1, S--Growth-limiting substrate concentration in culture broth, g.l-1, SR--Growth-limiting substrate concentration in feed, g.l-1, mean--Biomass concentration in culture broth, g.l-1, YS--Yield constant, g cell dry mass.g substrate-1, mu--Specific growth rate, h-1, mu max--Maximum growth rate, h-1.

Antonie Van Leeuwenhoek, 1990 Jan, 57(1), 29 - 32
Kinetic studies of phenol degradation by Rhodococcus sp . P1 . I . Batch cultivation; Straube G et al.; Rhodococcus sp . P1 utilizes phenol as the sole carbon and energy source via the beta-ketoadipate pathway . In batch cultivation, concentrations up to 2.8 g.l-1 phenol were degraded . The highest values for the specific growth rate of 0.32 h-1 were obtained at concentrations near 0.25 g.l-1 . At higher concentrations, substrate inhibition was observed, characterized by increases in lag phase and decreasing growth rates . A mathematical expression was proposed to fit the kinetic pattern of phenol inhibition on the specific growth rate mu: {formula: see text} Nomenclature: K- Exponent of the inhibition function, Ks- Monod saturation constant, g.l-1, KI- Inhibition constant, g.l-1, S- Substrate concentration in culture broth, g.l-1, So- Initial substrate concentration, g.l-1, Y- Yield constant, g cell dry mass.g substrate-1, mu- Specific growth rate, h-1, mu max- Maximum growth rate, h-1.

Scand J Infect Dis, 1990, 22(2), 227 - 32
Infected wounds and repeated septicemia in a case of factitious illness; Castor B et al.; During 19 years an assistant nurse, now 35 years old, has been repeatedly treated for several malingered and self-induced disorders escalating to self-mutilation . An ulcer of her right leg never epithelialised in spite of various local treatments and surgical intervention . During repeated attacks of self-induced septicemia altogether 11 different bacterial species were isolated; on 8 occasions Rhodococcus equi . The septicemias were successfully treated with antibiotics . The underlying psychiatric problem, a borderline personality disorder, has not been possible to treat in a conventional manner . Probably due to collaboration between the plastic surgeon and the psychiatrist she has had fewer attendances and shorter hospital stays lately . Her prognosis is still dubious as regards further self-mutilation and other expressions of self-destructive behaviour.

J Gen Microbiol, 1990 Jan, 136 ( Pt 1), 115 - 20
Isolation and characterization of a haloalkane halidohydrolase from Rhodococcus erythropolis Y2; Sallis PJ et al.; Rhodococcus erythropolis strain Y2, isolated from soil by enrichment culture using 1-chlorobutane, was able to utilize a range of halogenated aliphatic compounds as sole sources of carbon and energy . The ability to utilize 1-chlorobutane was conferred by a single halidohydrolase-type haloalkane dehalogenase . The presence of the single enzyme in cell-free extracts was demonstrated by activity strain polyacrylamide gel electrophoresis . The purified enzyme was a monomeric protein with a relative molecular mass of 34 kDa and demonstrated activity against a broad range of haloalkanes, haloalcohols and haloethers . The highest activity was found towards alpha, omega disubstituted chloro- and bromo- C2-C6 alkanes and 4-chlorobutanol . The Km value of the enzyme for 1-chlorobutane was 0.26 mM . A comparison of the R . erythropolis Y2 haloalkane halidohydrolase with other haloalkane dehalogenases is discussed on the basis of biochemical properties and N-terminal amino acid sequence data.

Pediatr Pathol, 1990, 10(3), 417 - 24
Pulmonary malacoplakia in a child; Byard RW et al.; Pulmonary malacoplakia is a rare condition with only five previously reported cases in the literature, all occurring in adults . We describe a case of malacoplakia of the lung in a 6-year-old female with acute lymphoblastic leukemia in remission and a Rhodococcus equi pneumonia and septicemia . The case demonstrates that pulmonary malacoplakia can occur in immunocompromised pediatric patients and that malacoplakia should, therefore, be added to the list of causes of nodular lung infiltrates in immunocompromised children.

Microbiol Immunol, 1990, 34(1), 45 - 53
Granuloma-forming activity and antitumor activity of newly isolated mycoloyl glycolipid from Rhodococcus terrae 70012 (Rt . GM-2); Natsuhara Y et al.; A newly isolated mycoloyl glycolipid (Rt . GM-2) from Rhodococcus terrae 70012 was identified and the granulomagenic and antitumor activities were studied as compared with trehalose-6,6'-dimycolate (cord factor) also from R . terrae (Rt . TDM) . The alkaline hydrolysis products of Rt . GM-2 contained trehalose, methyl-alpha-mycolate and a less-polar ester than the usual methyl-alpha-mycolate, possibly beta-keto mycolate (1:1:1, by mol . ratios) . On the other hand, analysis of alditol acetate obtained after the mild permethylation, NaBH4 reduction, and acetylation showed the occurrence of 2,3,4-tri-O-methyl-6-O-acetylglucitol . Therefore, the original glycolipid (Rt . GM-2) was identified tentatively as 6-O-alpha-mycoloyl 6'-O-beta-ketomycoloyl trehalose . Intravenous injection of Rt . GM-2 in the form of water-in-oil-in-water emulsion caused prominent granulomas in lungs and spleen of ICR and BALB/c mice . The granulomagenic effects were as strong as those caused by Rt . TDM . The lung and spleen weights reached peaks one week after an injection of Rt . GM-2 in mice and then gradually decreased . Multiple intravenous injections of Rt . GM-2 and Rt . TDM showed antitumor activity against subcutaneously implanted Sarcoma-180, and caused prominent granulomatous changes and growth suppression of mice.

Cancer Immunol Immunother, 1990, 31(2), 99 - 106
Parallel antitumor, granuloma-forming and tumor-necrosis-factor-priming activities of mycoloyl glycolipids from Nocardia rubra that differ in carbohydrate moiety: structure-activity relationships; Natsuhara Y et al.; Multiple intravenous injections (30 micrograms, ten times) in ICR mice of trehalose dimycolate and glucose monomycolate from Nocardia rubra, containing C36-48 mycolic acids, showed a prominent antitumor effect on a subcutaneously implanted sarcoma-180, an allogeneic sarcoma of mice with a significant granuloma formation in lungs, spleen and liver . On the other hand, mycoloyl glycolipids other than glucose monomycolate and trehalose dimycolate, such as mannose or fructose mycolate, showed no significant activity for tumor regression or granuloma formation in mice . Trehalose dimycolate and glucose monomycolate from N . rubra, and glucose monomycolate with C56-60 mycolic acids from Rhodococcus terrae also showed a distinctive priming activity for tumor necrosis factor (TNF), when lipopolysaccharide from Escherichia coli was administered as an eliciting agent . The TNF activity in the sera of mice was abrogated almost completely by anti-(murine TNF alpha) antibody with protein-A-agarose . Again in contrast, mannose and fructose mycolate from N . rubra and glucose monomycolate with C30-34 mycolic acids from Rhodococcus equi did not show such activities in mice . Meth-A, a syngeneic fibrosarcoma of BALB/c mice, was less sensitive to administration of glycolipids than sarcoma-180 . These results indicated that the existence of a glucose or trehalose molecule was necessary for the expression of immunomodifying activities among various mycoloyl glycolipids differing in carbohydrate structure . However, since the administration of lipopolysaccharide was essentially required as an eliciting agent for the induction of TNF, while no eliciting agent was required for the antitumor activities, TNF does not seem to contribute directly to the antitumor activities of mycoloyl glycolipids in our systems . There was, however, a parallel structure-activity relationship among granuloma-forming, antitumor and TNF-priming activities, indicating that the structures of both the carbohydrate moiety and the mycoloyl residues influenced an initial step, such as macrophage activation, commonly and profoundly.

Microbiol Immunol, 1990, 34(8), 697 - 701
Enhancement of natural resistance of mice to Rhodococcus equi infection by Dextran Sulfate 500; Takai S et al.; The effect of intraperitoneal injection of Dextran Sulfate 500 (DS-500, 500,000 Mr) on the intravenous clearance and host resistance to Rhodococcus equi in mice was studied . Contrary to the results seen with Listeria monocytogenes, DS-500 enhanced murine resistance to R . equi and altered organ distribution and clearance.

J Basic Microbiol, 1990, 30(6), 415 - 23
Steroid-1-dehydrogenases in nocardioform bacteria studied by electrophoresis and immuno blotting techniques; Kaufmann G et al.; Fifteen noncardioform bacteria strains, capable of transforming steroid compounds were investigated with regard to their range of inducible steroid-1-dehydrogenase (St1DH)1 activities . The St1DHs of these bacteria were compared due to their immuno reactivity in Western blot experiments with a rabbit antiserum raised against the purified St1DH of Rhodococcus rhodochrous 7030 . Four strains exhibited a strong immuno reactivity, irrespective of differences in the electrophoretic mobility of the enzymes . Five strains revealed significantly diminished reactivities, and in five strains with a very low St1DH content, no reactivity was found . One strain, designated as Nocardiaspec . 7151, exhibited a high, inducible St1DH activity, but no immunoreaction was found . The absence of immuno reactivity is discussed in connection with the considerably diminished electrophoretic mobility of this enzyme.

Microbiol Immunol, 1990, 34(6), 523 - 32
Induction of interferons (IFNs) and tumor necrosis factor (TNF) in mice by a novel glycolipid trehalose 2,3,6'-trimycolate from Rhodococcus aurantiacus (Gordona aurantiaca); Fujita T et al.; The immunomodifying activity of a novel mycoloyl glycolipid, trehalose 2,3,6'-trimycolate (GaGM), from a unique psychrophilic acid-fast bacterium, Rhodococcus aurantiacus, was examined . ICR mice were primed intravenously (i.v.) or intraperitoneally (i.p.) with liposomes containing GaGM (300 micrograms/mouse), and were administered LPS dissolved in saline (25 micrograms/mouse, i.v.) 2 weeks later . Two hours after injection of LPS, interferons (IFNs) and tumor necrosis factor (TNF) were induced significantly in mice sera . The increase in activities of IFNs and TNF was approximately paralleled with granuloma formation in spleen of mice primed with GaGM . However, IFNs and TNF were not induced either in mice primed with GaGM but not elicited with LPS, or in those primed with GaGM and elicited by GaGM . Both activities induced were lower in mice primed with trehalose mono- or dimycolate from R . aurantiacus (GaTMM, GaTDM) or TDM from Nocardia rubra than in GaGM-primed mice . Time course study showed that the maximum activity of each interferon (alpha, beta, or gamma) was observed at different stages after LPS administration; IFN-alpha, IFN-beta, and IFN-gamma appeared 3, 2, and 6 hours most abundantly after LPS administration, respectively.

Zentralbl Bakteriol, 1989 Dec, 272(2), 154 - 70
Lipid composition in the classification of Rhodococcus equi; Barton MD et al.; The fatty acid, menaquinone and polar lipid composition of representatives of Rhodococcus equi and related taxa were determined . All of the R . equi strains had major proportions of straight chain saturated, monounsaturated and 10-methyl branched fatty acids, dihydrogenated menaquinones with eight isoprene units as the predominant isoprenologue, and characteristic polar lipid patterns that contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides and glycolipids including a "cord factor"-like compound that was most pronounced in fresh isolates . The mycolic acids of these strains fell within the range C24 to C48, had 0 to 4 double bonds and released major amounts of C14:0 esters on pyrolysis . These lipid data provide further evidence that R . equi strains form a distinct taxospecies within the genus Rhodococcus . The remaining strains also gave lipid profiles consistent with their assignment to the genus Rhodococcus . These organisms included strains identified as R . sputi.

Infect Immun, 1989 Dec, 57(12), 3928 - 35
Purification and characterization of two Listeria ivanovii cytolysins, a sphingomyelinase C and a thiol-activated toxin (ivanolysin O); Vazquez-Boland JA et al.; The strong bizonal hemolysis on blood agar and the positive CAMP reaction with Rhodococcus equi denotes the production of two different cytolytic factors by Listeria ivanovii . One was characterized as a thiol-activated (SH) cytolysin of 61 kilodaltons and was termed ivanolysin O (ILO) since data suggested that it is different from listeriolysin O, the SH-cytolysin produced by Listeria monocytogenes . The other is a 27-kilodalton hemolytic sphingomyelinase C that was found to be the cytolytic factor responsible for the halo of incomplete hemolysis synergistically enhanced by R . equi exosubstances . When thiol-disulfide exchange affinity chromatography and gel filtration were applied to the purification of ILO from concentrated L . ivanovii culture supernatants, the copurification of the two cytolysins was observed . This phenomenon seems to be due to the formation of intermolecular disulfide bonds between ILO and the sphingomyelinase, since the latter was found to contain free SH groups, not essential for the activity . These SH groups could react with the single cysteine residue characteristically present in the SH-cytolysins, forming a dimeric cytolytic complex . The purification of ILO was achieved by a further gel filtration with a reducing agent (dithiothreitol) in the eluent . A method for the purification of the sphingomyelinase based on selective sequestration of ILO from the L . ivanovii concentrated culture supernatant by the SH cytolysin target molecule cholesterol and thiol-disulfide affinity chromatography is described.

Vet Immunol Immunopathol, 1989 Nov 15, 22(4), 369 - 78
The effect of immunosuppression on resistance to Rhodococcus equi in mice; Bowles PM et al.; Rhodococcus equi, a natural pathogen of horses, produces lesions in mice following experimental infection . The effect of various immunosuppressing agents on the sequential development of these lesions has been assessed by measuring the growth of R . equi following intravenous or intranasal challenge and by histological examination . Cyclophosphamide treatment of mice, challenged intranasally, resulted in the development of lesions not unlike that seen in experimental and natural infection in foals . Cortisone acetate also impaired bacterial clearance from the lungs and affected the accumulation of mononuclear cells at infective foci . Most of the agents chosen to impair macrophage function failed to affect the resistance of mice to R . equi . Carbon, carrageenan and silica failed to alter significantly the growth kinetics of R . equi . Dextran sulphate depressed the rate of pulmonary clearance of organisms and affected the ability of animals to eliminate R . equi following rechallenge . Overall, these results support other evidence that cell mediated immunity is involved in host resistance to R . equi and that activated macrophages play a role in acquired immunity to this organism.

Am J Kidney Dis, 1989 Nov, 14(5), 417 - 8
Rhodococcus peritonitis in a patient treated with peritoneal dialysis; Brown E et al.; This report is of a patient treated with continuous ambulatory peritoneal dialysis who developed peritonitis attributed to a Rhodococcus species . The rhodococci are rarely pathogenic to humans, and there are no previous reports of peritonitis associated with these organisms . It is important for clinicians to be aware of these bacteria because they can be acid-fast and can be confused with mycobacteria . They are often reported as "diphtheroids" but should not be considered contaminants.

J Comp Pathol, 1989 Nov, 101(4), 411 - 20
Early events associated with experimental infection of the murine lung with Rhodococcus equi; Bowles PM et al.; Pneumonia due to Rhodococcus equi was induced in the murine lung by deposition of a known dose of organisms . From serial estimations of bacterial numbers in the lungs of inoculated mice, analysis of the cellular composition of bronchoalveolar lavage fluid and morphological examination of the lungs, events in the host-parasite interaction were followed until day 7 . Early bacterial clearance from the lung was dose-dependent but was not sustained . A proportion of the inoculated R . equi was susceptible to the early nonspecific phagocytic cell response, and the contribution of neutrophils to bacterial clearance appeared largely limited to the first 24 hours . A substantial fraction of the organisms survived in the alveoli, probably within macrophages . The contribution phagocytes make to resistance against R . equi is similar to that which prevails in infection with Listeria monocytogenes.

Biochim Biophys Acta, 1989 Sep 11, 1005(1), 45 - 50
Composition and toxicity of lipids from Rhodococcus rhodochrous grown on medium containing galactose, glucose or mannose; de Almeida ET et al.; Rhodococcus rhodochrous, a producer of mycolic acid of approx . C40, exhibited a higher cellular mass yield when grown on glucose than when grown on galactose or mannose . The cellular content of the diethyl ether-soluble lipids in microorganisms cultivated on glucose or mannose varied with the incubation time, while that of microorganisms grown on galactose remained constant . The lipids extracts from cells cultivated on different hexoses and collected at the exponential phase of growth were more toxigenic; this property was related in general to the content of glycolipid . On the other hand, cells cultivated on galactose or mannose had a higher quantity of glycolipid in the exponential phase, while the glycolipid content of those grown on glucose remained approximately constant . Amongst the components of the lipid extract, the glycolipid fraction was the sole fraction bearing toxic property . Neutral plus fatty acids and phospholipids displayed no similar characteristic.

Steroids, 1989 Sep, 54(3), 271 - 86
Synthesis of 9 alpha-hydroxysteroids by a Rhodococcus sp; Datcheva VK et al.; 9 alpha-Hydroxylation of delta 5-3 beta-hydroxysteroids (of androstane, pregnane, 24-nor- and 21,24-bisnorcholane groups) was carried out by a Rhodococcus sp., isolated from a petroleum-containing soil sample . A large number of the investigated steroids was transformed into 9 alpha-hydroxy-delta 4-3-ketones in satisfactory yields (50-90%) at high initial concentrations of the substrates (0.5-5.0 g/L) . The influence of some structural features of the steroid molecule on the progress and effectiveness of the microbial transformation was also shown.

Biochem J, 1989 Aug 15, 262(1), 303 - 12
Purification and characterization of 4-methylmuconolactone methyl-isomerase, a novel enzyme of the modified 3-oxoadipate pathway in nocardioform actinomycetes; Bruce NC et al.; The novel enzyme 4-methyl-2-enelactone methyl-isomerase was detected in, and purified to electrophoretic homogeneity from, p-toluate-grown cells of Rhodococcus rhodocrous N75, a nocardioform actinomycete . The enzyme was very thermostable and had a native Mr of 75,500; as the monomer had an Mr of 17,000, the enzyme is probably tetrameric . The new isomerase is highly specific with respect to its lactone substrate, only accepting (+)-(4S)-4-methylmuconolactone (4-carboxymethyl-4-methylbut-2-en-1,4-olide), and the putative isomerization reaction intermediate 1-methylbislactone ((-)-1-methyl-3,7-dioxo-2,6-dioxabicyclo-{3.3.0}octane) as substrates, and yielding (-)-(4S)-3-methylmuconolactone (4-carboxymethyl-3-methylbut-2-en-1,4-olide) as product . Some other lactone analogues acted as competitive inhibitors . Our data suggest that the isomerization does not involve actual methyl migration, but proceeds via the 1-methybislactone.

Anal Biochem, 1989 Jul, 180(1), 91 - 4
Monitoring of phenylketonuria: a colorimetric method for the determination of plasma phenylalanine using L-phenylalanine dehydrogenase; Wendel U et al.; A simple, rapid, accurate, and precise colorimetric assay for the determination of L-phenylalanine in plasma samples using L-phenylalanine dehydrogenase {L-phenylalanine:NAD+-oxidoreductase (deaminating)} from Rhodococcus sp . M 4 is described . The enzyme catalyzes the NAD-dependent oxidative deamination of L-phenylalanine . However, the equilibrium of reaction favors L-phenylalanine formation . By stoichiometric coupling of this reaction with diaphorase/iodonitro tetrazolium chloride (INT) the formed NADH converts INT to a formazan whereby the reaction is displaced in favor of phenylpyruvate . Using a kinetic approach the increase in absorbance at 492 nm shows linearity over more than 30 min . Deproteinized standard solutions of L-phenylalanine in the range from 30 to 1200 mumol/liter show a linearity between the dAformazan/30 min and the substrate concentration . In phenylketonuria (PKU) plasma samples no interferences caused by L-tyrosine or phenylpyruvic acid are seen . Applicability is demonstrated by comparative determination of plasma L-phenylalanine of treated PKU patients by the colorimetric method and automated amino acid analysis.

Equine Vet J, 1989 Jul, 21(4), 249 - 55
Rhodococcus equi foal pneumonia: protective effects of immune plasma in experimentally infected foals; Martens RJ et al.; The immunoprophylactic capacity of specific immune plasma was evaluated in pony foals infected experimentally with Rhodococcus equi . Immune plasma, produced by repeated parenteral administration of viable R . equi to adult horses, was harvested and frozen . Group I (six control foals) and Group II (six principal foals) received lactated Ringers solution and immune plasma respectively at three and five days of age . R . equi were aerosolised into a caudal lung lobe of all foals at seven days of age . Clinical signs, haematological alterations, immune responses, thoracic radiographs and technetium99m pulmonary perfusion scans were monitored . All foals were destroyed and complete post mortem examinations performed . All foals developed pneumonia as evidenced by clinical, radiographic and perfusion alterations, but the survival rate of principal foals was significantly (P less than 0.01) greater than that of control foals . Five control foals developed terminal disease, whereas all principal foals recovered . There was no significant (P greater than 0.05) difference in temperature response, or peripheral blood leucocyte, neutrophil or fibrinogen concentrations between groups . ELISA values for R . equi antibody were significantly (P less than 0.001) greater in principal foals following treatment, but there was no significant (P greater than 0.05) difference in IgG or IgM concentrations between groups . Results of the haemolysis inhibition assay indicated that equi factor neutralising antibodies were transferred by immune plasma to the principal foals . Post mortem examinations of five control foals destroyed at approximately three weeks post infection because of terminal disease, revealed severe pyogranulomatous pneumonia . One control and all principal foals were either free of lesions or had resolving lesions and/or minimal scar formation at three months post infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Biochem, 1989 Jun 15, 182(2), 349 - 56
Nitrilase of Rhodococcus rhodochrous J1 . Purification and characterization; Kobayashi M et al.; Nitrilase was purified from an extract of isovaleronitrile-induced cells of Rhodococcus rhodochrous J1 in seven steps . In the last step, the enzyme was crystallized by adding ammonium sulfate . The crystallized enzyme appeared to be homogeneous by polyacrylamide electrophoresis, ampholyte electrofocusing and double immunodiffusion in agarose . The enzyme has a molecular mass of about 78 kDa and consists of two subunits identical in molecular mass . The purified enzyme exhibits a pH optimum of 7.6 and a temperature optimum of 45 degrees C . The enzyme catalyzed stoichiometrically the hydrolysis of benzonitrile to benzoic acid and ammonia, and no formation of amide was detected . The enzyme required thiol compounds such as dithiothreitol, L-cysteine or reduced glutathione to exhibit maximum activity . The enzyme was specific for nitrile groups attached to an aromatic or heteroaromatic ring, e.g . benzonitrile, 3-chlorobenzonitrile, 4-tolunitrile, 2-furonitrile and 2-thiophenecarbonitrile . The comparison of the properties of the enzyme with other nitrilases and nitrile hydratases has been also discussed.

J Biol Chem, 1989 Jun 5, 264(16), 9510 - 9
Purification and characterization of glycosphingolipid-specific endoglycosidases (endoglycoceramidases) from a mutant strain of Rhodococcus sp . Evidence for three molecular species of endoglycoceramidase with different specificities; Ito M et al.; Two molecular species of endoglycoceramidase (designated as endoglycoceramidases I and II) were purified 32,700 and 43,000 times with overall recoveries of 4.8 and 2.9%, respectively, from a culture fluid of the mutant strain M-750 of Rhodococcus sp., cultivated in the absence of inducers (ganglioside) . After being stained with Coomassie Brilliant Blue or a silver-staining solution, each purified enzyme showed a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate . The apparent molecular weights, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were 55,900 for endoglycoceramidase I and 58,900 for endoglycoceramidase II, and their pIs were 5.3 and 4.5, respectively . both were capable of hydrolyzing the glucosylceramide linkage of ganglio-type, lacto-type, and globo-type glycosphingolipids to afford intact oligosaccharides and ceramides . Globo-type glycosphingolipids were strongly resistant to hydrolysis by endoglycoceramidase II in comparison with endoglycoceramidase I . Neither could hydrolyze gala-type glycosphingolipids, cerebrosides, sulfatides, glycoglycerolipids, or sphingomyelins . In addition to these two enzymes, the strain M-750 produced a third minor molecular species of endoglycoceramidase designated as endoglycoceramidase III . It was found capable of specifically hydrolyzing the galactosylceramide linkage of gala-type glycosphingolipids that were not hydrolyzable at all by endoglycoceramidases I or II . The molecular weights of the oligosaccharide and ceramide released from asialo GM1, incubated either in normal H2O or H2(18)O with the enzyme, were compared by fast atom bombardment-mass spectrometry . The result clearly indicated that both endoglycoceramidases I and II hydrolyze the glycosidic linkage between the oligosaccharide and ceramide . Thus, a systematic name of the endoglycoceramidase should be glycosyl-N-acyl-sphingosine 1,1-beta-D-glucanohydrolase.

J Appl Bacteriol, 1989 Jun, 66(6), 497 - 506
Evidence for the pathogenic role of Rhodococcus species in pulmonary diseases; Osoagbaka OU; Three hundred expectorated sputa from patients suffering from chest disorders were examined microscopically and cultured on various media . Among other micro-organisms isolated there were four strains of Rhodococcus aurantiacus (Gordona aurantiaca), one each of Rh . erythropolis, Rh . pellegrino, Rh . rubropertinctus and Rh . rhodnii . Suspensions in 5% hog gastric mucin were virulent for immuno-suppressed white mice and the pathology of their lungs was compatible with those produced in experimental nocardial and rhodococcus infections . The in-vitro antibiogram was similar to that of Nocardia asteroides . Two of the patients from whom Rh . aurantiacus and Rh . pellegrino were isolated responded bacteriologically and clinically to treatment with co-trimoxazole . The frequency with which these rhodococci were isolated and the clinical conditions of the patients strongly indicated a pathogenic role for some Rhodococcus species . It is suggested that rhodococci should be sought in chronic pulmonary infections, particularly in those who are immuno-compromised or debilitated, and their aetiologic role determined.

J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1507 - 13
Cloning and expression of Rhodococcus genes encoding pigment production in Escherichia coli; Hill R et al.; Pigment was produced by Escherichia coli cells carrying recombinant plasmids pNIL100, pNIL200 and pNIL400 containing DNA from Rhodococcus sp . E . coli cells containing pNIL100 or pNIL200 (with DNA inserts from Rhodococcus sp . JL10 and Rhodococcus sp . ATCC 21145 respectively) produced both blue and pink pigments, while cells containing pNIL400 (with a DNA insert from Rhodococcus sp . ATCC 21145) produced only pink pigment . Colonies of E . coli(pNIL100) and E . coli(pNIL200) were dark blue, whereas E . coli(pNIL400) colonies were pink . No pigment was detected in Streptomyces griseus transformants containing pNIL100, pNIL200 or pNIL400 . Restriction endonuclease mapping indicated that the cloned DNA fragments were different . The pigment gene(s) in pNIL200 producing both the blue and pink pigments were contained within a 2.8 kb DNA fragment . The pigments produced by E . coli transformants containing pNIL200 were characterized by visible and UV spectroscopy . No similar pigments were detected in Rhodococcus sp . ATCC 21145.

Eur J Biochem, 1989 May 15, 181(3), 563 - 70
Primary structure of nitrile hydratase deduced from the nucleotide sequence of a Rhodococcus species and its expression in Escherichia coli; Ikehata O et al.; The nitrile hydratase (NHase) of Rhodococcus species N-774, which is composed of two subunits, alpha and beta, catalyzes the hydration of various nitrile compounds to the corresponding amides . The amino acid sequences of the NH2 termini and the fragments obtained by digesting each of the two subunits with lysyl endopeptidase were determined for preparation of synthetic oligonucleotides as hybridization probes . A 4.4-kb SphI fragment which contained DNA sequences hybridizing to several of the probes was cloned in pBR322 in Escherichia coli . The nucleotide sequences together with the determined amino acid sequences indicated that the alpha and beta subunits of NHase consisted of 207 amino acids (Mr, 22918) and 212 amino acids (Mr, 23428), respectively . The open reading frame for the alpha subunit includes that for the beta subunit with a short interval of only 26 base pairs; the two genes are probably translated in a polycistronic manner . Although large amounts of the alpha- and beta-subunit proteins were produced as insoluble forms in E . coli when the cloned genes were placed under the control of the lac promoter, no enzymatic activity was detected . The activity of the enzyme was restored, to some extent, by solubilization of the proteins with 8 M urea and subsequent dialysis for refolding at pH 10 in the presence of Fe2+ and pyrroloquinoline quinone.

Schweiz Med Wochenschr, 1989 May 6, 119(18), 566 - 74
{Rhodococcus equi infection in HIV disease}; Flepp M et al.; A case of Rhodococcus equi pneumonia associated with septicemia and metastatic brain abscess in an HIV-infected male is presented . Clinical findings and the diagnostic and therapeutic approach are discussed . Cavitating pulmonary disease was rapidly improved by six-week combined oral and parenteral antibiotic treatment . However, pulmonary relapse and brain abscess were documented after one month . Therapy with ceftriaxone, ciprofloxacin and cotrimoxazole on an outpatient basis again led to clinical and radiological improvement after four weeks . Surgical resection of localized processes should be considered early, and prolonged antibiotic therapy over months is recommended.

Aust N Z J Med, 1989 Apr, 19(2), 103 - 7
Rhodococcus equi: an emerging opportunistic pathogen?
Jones MR, Neale TJ, Say PJ, Horne JG.
Human infection with Rhodococcus equi is apparently rare with most published reports describing the development of lung abscesses in immunocompromised hosts . Of only 18 cases of infection previously recorded, four have recently occurred in patients with the acquired immune deficiency syndrome (AIDS) . In Australasia, R . equi has frequently been isolated from soil and infected farm animals yet no human infections have been reported thus far . Three cases of R . equi infection have occurred in New Zealand and, collectively, they cover a wider spectrum of disease than that previously recognised . The natural history of R . equi infections, their clinical features and treatment are described in the light of our recent experience.

FEBS Lett, 1989 Jan 16, 243(1), 61 - 4
Nitrile hydratase of Rhodococcus sp . N-774 . Purification and amino acid sequences; Endo T et al.; The nitrile hydratase of Rhodococcus sp . N-774 was purified and crystallized . The enzyme is composed of two different subunits (molecular masses: subunit alpha, 28,500 Da; subunit beta, 29,000 Da) . The amino-terminal amino acid sequence of each subunit was determined . There is no sequence homology between the two subunits, suggesting that the peptides originate from different cistrons . The activity of the purified enzyme did not decrease during incubation in the dark, whereas it gradually decreased in intact cells.

Microbiol Immunol, 1989, 33(6), 503 - 8
Carotenoid pigments of genus Rhodococcus; Ichiyama S et al.; A study of carotenoid pigments of the genus Rhodococcus was carried out . According to carotenes contained, Rhodococcus species were divided into three groups: the first group of Rhodococcus luteus, R . coprophilus, R . lentifragmentus, and R . maris, which formed beta-carotene; the second group of R . equi, R . rubropertinctus, R . aichiensis, R . sputi, R . chubuensis, R . obuensis, R . bronchialis, R . roseus, R . rhodochrous, R . rhodnii, and R . terrae, which formed gamma-carotene-like substance; and the third group of R . aurantiacus, which formed neither carotene . Other carotenoid pigments were different according to the species.

Yakugaku Zasshi, 1989 Jan, 109(1), 46 - 51
{Granuloma formation in mice by liposomes of new glycolipid containing mycolic acids, "trehalose 2,3,6'-trimycolate"}; Yokoi F et al.; After intravenous administration of liposomes containing a novel glycolipid, trehalose 2,3,6'-trimycolate, isolated from Rhodococcus aurantiacus, the granuloma formation was observed distinctively in the lungs, spleen and liver of ICR mice . The concentration of the glycolipid in liposomes influenced profoundly on the inducibility for granulomas and the liposomes containing 10 mol% of trehalose 2,3,6'-trimycolate showed the highest activity for the granuloma formation in mice without a significant loss of body weight of mice with a less toxicity . EggPC liposomes are of more highly inducible for the granuloma formation in mice than eggPC:PS (7:3, mol ratio) liposomes or eggPC:Chol (9:1, mol ratio) liposomes, suggesting that cholesterol in liposomes was not necessarily essential for inducing granulomas in mice . Among the various PC liposomes, synthetic dimyristoyl-phosphatidylcholine (DMPC) or dipalmitoylphosphatidylcholine (DPPC) liposomes were of highly inducible, although a high responsiveness was also observed with natural lecithines such as egg or soy PC . However, distearoylphosphatidylcholine (DSPC) liposomes did not show any activity for the granuloma formation in any organs of mice . Organ responsiveness differed significantly in the micelle forms injected . Glycolipid entrapped with eggPC-Chol liposomes showed a lower lung index below 1.0, in contrast to w/o/w micelles containing Freunds' incomplete adjuvant, which showed a higher granuloma formation in the lungs . It was also noted that the toxicity of glycolipid containing liposomes was comparatively lower than the toxicity of glycolipid containing w/o/w micelles, indicating that the liposomes appeared to be more suitable for the induction of the immunomodifying activities of mycolic acid-containing glycolipids than w/o/w micelles.

J Bacteriol, 1989 Jan, 171(1), 30 - 6
Purification and characterization of a dimeric phenylalanine dehydrogenase from Rhodococcus maris K-18; Misono H et al.; NAD+-dependent phenylalanine dehydrogenase (EC 1.4.1.) was purified to homogeneity from a crude extract of Rhodococcus maris K-18 isolated from soil . The enzyme had a molecular mass of about 70,000 daltons and consisted of two identical subunits . The enzyme catalyzed the oxidative deamination of L-phenylalanine and several other L-amino acids and the reductive amination of phenylpyruvate and p-hydroxyphenylpyruvate . The enzyme required NAD+ as a natural coenzyme . The NAD+ analog 3-acetylpyridine-NAD+ showed much greater coenzyme activity than did NAD+ . D-Phenylalanine, D-tyrosine, and phenylethylamine inhibited the oxidative deamination of L-phenylalanine . The enzyme reaction was inhibited by p-chloromercuribenzoate and HgCl2 . Initial-velocity and product inhibition studies showed that the reductive amination proceeded through a sequential ordered ternary-binary mechanism . NADH bound first to the enzyme, followed by phenylpyruvate and then ammonia, and the products were released in the order L-phenylalanine and NAD+ . The Michaelis constants were as follows: L-phenylalanine, 3.8 mM; NAD+, 0.25 mM; NADH, 43 microM; phenylpyruvate, 0.50 mM; and ammonia, 70 mM.

Appl Environ Microbiol, 1988 Dec, 54(12), 3043 - 52
Degradation and O-methylation of chlorinated phenolic compounds by Rhodococcus and Mycobacterium strains; Haggblom MM et al.; Three polychlorophenol-degrading Rhodococcus and Mycobacterium strains were isolated independently from soil contaminated with chlorophenol wood preservative and from sludge of a wastewater treatment facility of a kraft pulp bleaching plant . Rhodococcus sp . strain CG-1 and Mycobacterium sp . strain CG-2, isolated from tetrachloroguaiacol enrichment, and Rhodococcus sp . strain CP-2, isolated from pentachlorophenol enrichment, mineralized pentachlorophenol and degraded several other polychlorinated phenols, guaiacols (2-methoxyphenols), and syringols (2,6-dimethoxyphenols) at micromolar concentrations and were sensitive to the toxic effects of pentachlorophenol . All three strains initiated degradation of the chlorophenols by para-hydroxylation, producing chlorinated para-hydroquinones, which were then further degraded . Parallel to degradation, strains CG-1, CG-2, and CP-2 also O-methylated nearly all chlorinated phenols, guaiacols, syringols, and hydroquinones . O-methylation of chlorophenols was a slow reaction compared with degradation . The preferred substrates of the O-methylating enzyme(s) were those with the hydroxyl group flanked by two chlorine substituents . O-methylation was constitutively expressed, whereas degradation of chlorinated phenolic compounds was inducible.

Res Vet Sci, 1988 Nov, 45(3), 416 - 7
Use of a monoclonal antibody in the diagnosis of infection by Dermatophilus congolensis; How SJ et al.; A monoclonal antibody (McAb) to Dermatophilus congolensis was produced from murine hybridoma cultures and purified by affinity chromatography . Species specificity was demonstrated using indirect immunofluorescent staining; the McAb was shown to react with 10 D congolensis isolates but not with 10 Nocardia species isolates, a Rhodococcus and a Streptomyces species isolate . The McAb was used to demonstrate D congolensis in clinical material from confirmed bovine and ovine cases and presumptive equine cases of dermatophilosis by indirect immunofluorescent staining.

J Gen Microbiol, 1988 Oct, 134 ( Pt 10), 2807 - 13
DNA base composition and homology values in the classification of some Rhodococcus species; Zakrzewska-Czerwinska J et al.; Similar DNA homology values were recorded when a modified S1 nuclease technique and a standard nitrocellulose membrane filter method were applied to representative strains of Rhodococcus . The DNA homology data showed that R . globerulus, R . luteus and R . sputi form distinct genomic species . The congruence between the DNA homology and earlier numerical phenetic data was good, but there was evidence that some strains had been misclassified in the previous studies . In particular, the type strains of R . obuensis and R . sputi belong to a single genomic species . The former name is thus a later, subjective synonym of the latter . The guanine plus cytosine content of the DNA of the rhodococci fell within the range 61 to 72 mol%.

Aust N Z J Med, 1988 Oct, 18(6), 790 - 1
Lung infection caused by Rhodococcus; Hart DH et al.; We report a case of lung infection, clinically resembling tuberculosis, caused by Rhodococcus rubropertinctus . The patient had no apparent immunosuppression which is unusual for disease caused by the 'rhodochrous' complex . The infection responded successfully to oral anti-tuberculous therapy, which included rifampicin, and to oral tetracycline.

Chest, 1988 Jul, 94(1), 195 - 6
Rhodococcus equi pneumonia . An unusual early manifestation of the acquired immunodeficiency syndrome (AIDS); Weingarten JS et al.; Infection with Rhodococcus equi has been reported as an occasional cause of cavitary pneumonia in severely immunocompromised patients, including those with the acquired immunodeficiency syndrome (AIDS) . We report two cases of R equi pneumonia presenting in one month in patients infected with human immunodeficiency virus (HIV) who had not previously had an opportunistic infection . The clinical and radiographic manifestations of the disease are distinctive and should suggest the diagnosis . R equi pneumonia in a person with HIV infection should be considered diagnostic of AIDS . Recognition of this entity is important since antibiotic therapy is different from that conventionally used in pneumonias in AIDS patients and must be prolonged.

Diagn Microbiol Infect Dis, 1988 Jul, 10(3), 185 - 90
Isolation of Rhodococcus rhodochrous from a chronic corneal ulcer; Gopaul D et al.; Organisms belonging to Rhodococcus species have been isolated as the causative agent of infections in many animals and humans . The majority of the human infections so far reported have been limited to immunocompromised patients including AIDS patients . We report an elderly woman with a chronic corneal ulcer infected with Rhodococcus . To our knowledge there is no previous report on a Rhodococcus infection of the eye . Rarity of this type of infection by Rhodococcus in a locally immunocompromised site in an otherwise healthy individual prompted this case history.

J Clin Microbiol, 1988 Jun, 26(6), 1221 - 2
Endophthalmitis caused by Rhodococcus equi Prescott serotype 4; Ebersole LL et al.; Rhodococcus equi, an aerobic actinomycete primarily known as a livestock pathogen, was repeatedly isolated from the severely infected right eye of a 9-year-old boy.

J Chromatogr, 1988 May 25, 440, 479 - 86
Investigation of the lipids of saprophytic mycobacteria in the U.S.S.R; Koronelli TV; Three trends in the investigations of the specific lipids of rhodococci and related microorganisms are reflected: isolation and structural determination of new complex lipids, elucidation of the role of specific lipids in the cells and application of lipid composition for diagnostic studies of rhodococci and related organisms . Two groups of peptidolipids, differing in chromatographic mobility and peptide chain structure, have been found in the cells of Rhodococcus erythropolis . The compounds within each group differ in acyl moieties . Three peptidolipids of the polar group include glucose, so they are peptidoglycolipids . Glycolipids are represented by trehalose derivates . Trehalose dimycolate (cord factor) is dominant . Data concerning the role of lipids in typical features of rhodococci as the lipophilic cell wall, the ability to up-take a hydrophobic substrate and the resistance to influences from outside are given . Examples of application of mycolic acid composition in taxonomy of rhodococci and morphologically similar bacteria are given.

J Bacteriol, 1988 May, 170(5), 2401 - 5
Conjugative transfer of cadmium resistance plasmids in Rhodococcus fascians strains; Desomer J et al.; The presence of a 138-kilobase plasmid (pD188) correlated with increased resistance to cadmium in Rhodococcus fascians D188 . This plasmid could be transferred by a conjugation-like system in matings between R . fascians strains . Transconjugants expressed the cadmium resistance and could be used as donors in subsequent matings . Four other R . fascians strains (NCPPB 1488, NCPPB 1675, NCPPB 2551, and ATCC 12974) could also be used as donors for cadmium resistance in matings . Strain NCPPB 1675 showed a 100% cotransfer of cadmium and chloramphenicol resistance markers.

Infection, 1988 May-Jun, 16(3), 186 - 8
Osteomyelitis caused by Rhodococcus equi in a renal transplant recipient; Novak RM et al.; We report the first case of osteomyelitis due to Rhodococcus equi, which occurred in a renal transplant patient . Infection with this organism is rare and usually causes a distinct clinical syndrome resembling pulmonary tuberculosis . We investigated by time-kill curve analysis various antimicrobial combinations for in vitro efficacy . The literature is briefly reviewed, and aspects of diagnosis and therapy are discussed.

J Clin Microbiol, 1988 Apr, 26(4), 618 - 20
Characterization of Rhodococcus equi-like bacterium isolated from a wound infection in a noncompromised host; Muller F et al.; A case of superficial wound infection in a noncompromised host due to a Rhodococcus equi-like organism, acquired from soil, is described . The strain grew poorly at 37 degrees C and showed several differences from wild-type R . equi in biochemical tests and in its fatty acid and mycolic acid compositions.

Can J Microbiol, 1988 Mar, 34(3), 242 - 8
Metabolism of acetylene and acetaldehyde by Rhodococcus rhodochrous; Germon JC et al.; We studied the ability of a soil bacterium, identified as Rhodococcus rhodochrous, to grow on acetylene and to accumulate acetaldehyde . Its maximum growth rate on acetylene was obtained at about 30 degrees C (mu = 0.11 h-1) and was independent of the concentration of this gas in air from 0.14 to 16% (v/v) . During growth, acetylene was quantitatively transformed to acetaldehyde, ethanol, acetate, CO2, and biomass in proportions which varied with culture age and temperature . Growth was completely inhibited by acetaldehyde at a concentration of 10 mM . The inhibitory effect was relieved by addition of acetate . Growth on ethanol up to 140 mM did not result in acetaldehyde accumulation . Acetylene consumption was constitutive with apparent Km and Vmax equal to 250 microM and 800 nmol.min-1.(mg protein)-1, respectively . In resting cell suspensions, acetylene consumption rates decreased more rapidly under air than under nitrogen . The inhibitory effect of acetaldehyde was enhanced in the presence of oxygen . Acetaldehyde accumulation in aerobic resting cell conditions did not exceed 10 mM (440 mg/L), but under anaerobic conditions it attained more than 70 mM (3.08 g/L).

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