Appl Environ Microbiol, 1999 Aug, 65(8), 3493 - 501 IS1631 occurrence in Bradyrhizobium japonicum highly reiterated sequence-possessing strains with high copy numbers of repeated sequences RSalpha and RSbeta; Isawa T et al.; From Bradyrhizobium japonicum highly reiterated sequence-possessing (HRS) strains indigenous to Niigata and Tokachi in Japan with high copy numbers of the repeated sequences RSalpha and RSbeta (K . Minamisawa, T . Isawa, Y . Nakatsuka, and N . Ichikawa, Appl . Environ . Microbiol . 64:1845-1851, 1998), several insertion sequence (IS)-like elements were isolated by using the formation of DNA duplexes by denaturation and renaturation of total DNA, followed by treatment with S1 nuclease . Most of these sequences showed structural features of bacterial IS elements, terminal inverted repeats, and homology with known IS elements and transposase genes . HRS and non-HRS strains of B . japonicum differed markedly in the profiles obtained after hybridization with all the elements tested . In particular, HRS strains of B . japonicum contained many copies of IS1631, whereas non-HRS strains completely lacked this element . This association remained true even when many field isolates of B . japonicum were examined . Consequently, IS1631 occurrence was well correlated with B . japonicum HRS strains possessing high copy numbers of the repeated sequence RSalpha or RSbeta . DNA sequence analysis indicated that IS1631 is 2,712 bp long . In addition, IS1631 belongs to the IS21 family, as evidenced by its two open reading frames, which encode putative proteins homologous to IstA and IstB of IS21, and its terminal inverted repeat sequences with multiple short repeats. Appl Environ Microbiol, 1999 Aug, 65(8), 3312 - 8 Characterization of methanotrophic bacterial populations in soils showing atmospheric methane uptake; Holmes AJ et al.; The global methane cycle includes both terrestrial and atmospheric processes and may contribute to feedback regulation of the climate . Most oxic soils are a net sink for methane, and these soils consume approximately 20 to 60 Tg of methane per year . The soil sink for atmospheric methane is microbially mediated and sensitive to disturbance . A decrease in the capacity of this sink may have contributed to the approximately 1% . year(-1) increase in the atmospheric methane level in this century . The organisms responsible for methane uptake by soils (the atmospheric methane sink) are not known, and factors that influence the activity of these organisms are poorly understood . In this study the soil methane-oxidizing population was characterized by both labelling soil microbiota with (14)CH(4) and analyzing a total soil monooxygenase gene library . Comparative analyses of {(14)C}phospholipid ester-linked fatty acid profiles performed with representative methane-oxidizing bacteria revealed that the soil sink for atmospheric methane consists of an unknown group of methanotrophic bacteria that exhibit some similarity to type II methanotrophs . An analysis of monooxygenase gene libraries from the same soil samples indicated that an unknown group of bacteria belonging to the alpha subclass of the class Proteobacteria was present; these organisms were only distantly related to extant methane-oxidizing strains . Studies on factors that affect the activity, population dynamics, and contribution to global methane flux of "atmospheric methane oxidizers" should be greatly facilitated by use of biomarkers identified in this study. J Vet Med Sci, 1999 Jun, 61(6), 717 - 20 Sequence analysis and expression of Nramp-1 gene in Bcgr and Bcgs mice; Nakanaga K et al.; Nucleic acid sequence of complemental DNA open reading frame for Nramp-1 gene was compared among DBA/2 (Bcgr), C57BL/6 (Bcgs) and C57BL/6-Bcgr mice which was previously developed as M . avium-resistant mouse strain (Xu, et al . Veterinary Microbiology 50:73-79 (1996) . Total RNA was isolated from various organs of DBA/2, C57BL/6 and C57BL/6-Bcgr . Nramp-1 cDNA was constructed from their mRNAs by gene amplification (PCR) technique and their open reading frame sequences were compared . The results clearly showed that our C57BL/6-Bcgr was almost identical with the DNA sequence of the DBA/2 mice . In contrast, C57BL/6-Bcgs mice differed only on the substitution of adenine for guanine of the nucleic acid at position 596 . This corresponded to the site of amino acid substitution (glycine to asparate) at position 169 in predicted NRAMP which had been reported . The expression of Nramp-1 mRNA was more prominent in spleens and livers and there appeared to be no significant difference among the strains of mice. Antonie Van Leeuwenhoek, 1999 Jan-Feb, 75(1-2), 5 - 19 The beta-lactam antibiotics: past, present, and future; Demain AL et al.; The discovery and development of the beta-lactam antibiotics are among the most powerful and successful achievements of modern science and technology . Since Fleming's accidental discovery of the penicillin-producing mold, seventy years of steady progress has followed, and today the beta-lactam group of compounds are the most successful example of natural product application and chemotherapy . Following on the heels of penicillin production by Penicillium chrysogenum came the discoveries of cephalosporin formation by Cephalosporium acremonium, cephamycin, clavam and carbapenem production by actinomycetes, and monocyclic beta-lactam production by actinomycetes and unicellular bacteria . Each one of these groups has yielded medically-useful products and has contributed to the reduction of pain and suffering of people throughout the world . Research on the microbiology, biochemistry, genetics and chemistry of these compounds have continued up to the present with major contributions being made by both individual and collaborative groups from industry and academia . The discovery of penicillin not only led to the era of the wonder drugs but provided the most important antibiotics available to medicine . Continued efforts have resulted in the improvement of these compounds with respect to potency, breadth of spectrum, activity against resistant pathogens, stability and pharmacokinetic properties . On the research front, major advances are being made on structural and regulatory biosynthetic genes and metabolic engineering of the pathways involved . New semisynthetic compounds especially those designed to combat resistance development are being examined in the clinic, and unusual non-antibiotic activities of these compounds are being pursued . Although seventy years of age, the beta-lactams are not yet ready for retirement. Ir J Med Sci, 1999 Apr-Jun, 168(2), 111 - 3 HpSA: assessment of a new non-invasive diagnostic assay for Helicobacter pylori infection in an Irish population; McNamara D et al.; BACKGROUND: The diagnosis of Helicobacter pylori is an essential element in the management of many common gastrointestinal pathologies . Previously diagnosis was dependent on the availability of endoscopic biopsy samples . The advent of non invasive assays such as the C13Urea breath test and Elisa serology have enabled diagnosis and treatment to be undertaken in the primary care setting . The isolation of Helicobacter pylori antigen from stool has led to the development of a new non-invasive test . AIM: A prospective study was designed to assess and compare the performance of Premier Platinum HpSA with current gold standard tests . METHODS: Consecutive patients undergoing a gastroscopy for investigation of dyspepsia at the Meath and Adelaide hospitals were enrolled . At endoscopy gastric biopsies were taken for histology, microbiology and rapid urease testing . In addition all subjects had C13UBT, serology and stool tests performed . Individuals who were H . pylori positive received standard proton pump inhibitor based triple therapy . Following treatment all tests, apart from serology were repeated . RESULTS: 54 patients were enrolled, 46 per cent were H . pylori positive . HpSA had a sensitivity and specificity and positive and negative predicted values of 96 per cent, 75 per cent and 80.6 per cent, 75.8 per cent respectively and compared favourably with all other tests . The sensitivity and specificities of the other tests were, histology 79.2 per cent and 100 per cent, culture 68 per cent and 100 per cent, rapid urease test 75 per cent and 100 per cent, serology 75 per cent and 96 per cent and C13 urea breath test 100 per cent and 96.6 per cent . CONCLUSION: The detection of H . pylori antigen in stool by means of a HpSA assay is a new and effective non-invasive means of diagnosis which can be performed in a routine laboratory setting . It is simple to perform and has possible advantages over other non-invasive tests, detecting actual antigen indicating current active infection. Cas Lek Cesk, 1999 May 24, 138(11), 329 - 32 {Lyme borreliosis}; Mrazek V et al.; The authors present a review of contemporary views on the most frequent anthropozoonosis--Lyme borreliosis . They mention its history, microbiology of the causal agent Borrelia burgdorferi, epidemiology, pathogenesis, laboratory and clinical diagnosis . The authors describe also briefly the clinical stages, forms and possible treatment. Plant J, 1999 Jun, 18(5), 571 - 6 A bacterial haloalkane dehalogenase gene as a negative selectable marker in Arabidopsis; Naested H et al.; The dhlA gene of Xanthobacter autotrophicus GJ10 encodes a dehalogenase which hydrolyzes dihalo- alkanes, such as 1, 2-dichloroethane (DCE), to a halogenated alcohol and an inorganic halide (Janssen et al . 1994, Annu . Rev . Microbiol . 48, 163-191) . In Xanthobacter, these alcohols are further catabolized by alcohol and aldehyde dehydrogenase activities, and by the product of the dhlB gene to a second halide and a hydroxyacid . The intermediate halogenated alcohols and, in particular, the aldehydes are more toxic than the haloalkane substrates or the pathway products . We show here that plants, including Arabidopsis, tobacco, oil seed rape and rice, do not express detectable haloalkane dehalogenase activities, and that wild-type Arabidopsis grows in the presence of DCE . In contrast, DCE applied as a volatile can be used to select on plates or in soil transgenic Arabidopsis which express dhlA . The dhlA marker therefore provides haloalkane dehalogenase reporter activity and substrate dependent negative selection in transgenic plants. Blood Rev, 1999 Jun, 13(2), 91 - 104 Microbiology confirmatory tests for blood donors; Dow BC; Blood donations collected in Scotland are currently screened for the presence of HBsAg, anti-HIV 1 + 2, anti-HCV and syphilis antibodies . Approximately 1% of all donations are found to repeatedly react to one of these four markers on screening but very few represent true infection . These samples must be sent to the designated confirmatory laboratory whose main role is to identify the true positive amongst a sea of 'false positives' . A battery of tests is used for this purpose, usually applied in a defined sequence . The use of such 'confirmatory algorithms' for each marker has been developed by most countries over the years and is now essential to the confirmatory process . The advent of gene amplification techniques such as PCR for initially pooled and eventually single donation testing will be the next challenge for confirmatory laboratories and will demand standards of confirmation as accurate as currently performed with the present serological markers. Rev Inst Med Trop Sao Paulo, 1999 Mar-Apr, 41(2), 87 - 94 Influence of microbiota in experimental cutaneous leishmaniasis in Swiss mice; de Oliveira MR et al.; Infection of Swiss/NIH mice with Leishmania major was compared with infection in isogenic resistant C57BL/6 and susceptible BALB/c mice . Swiss/NIH mice showed self-controlled lesions in the injected foot pad . The production of high levels of interferon-gamma (IFN-gamma) and low levels of interleukin-4 (IL-4) by cells from these animals suggests that they mount a Th1-type immune response . The importance of the indigenous microbiota on the development of murine leishmaniasis was investigated by infecting germfree Swiss/NIH in the hind footpad with L . major and conventionalizing after 3 weeks of infection . Lesions from conventionalized Swiss/NIH mice were significantly larger than conventional mice . Histopathological analysis of lesions from conventionalized animals showed abscesses of variable shapes and sizes and high numbers of parasitized macrophages . In the lesions from conventional mice, besides the absence of abscess formation, parasites were rarely observed . On the other hand, cells from conventional and conventionalized mice produced similar Th1-type response characterized by high levels of IFN-gamma and low levels of IL-4 . In this study, we demonstrated that Swiss/NIH mice are resistant to L . major infection and that the absence of the normal microbiota at the beginning of infection significantly influenced the lesion size and the inflammatory response at the site of infection. Curr Opin Rheumatol, 1999 Jul, 11(4), 244 - 50 Enthesitis in spondyloarthropathy; McGonagle D et al.; Inflammation at the insertions of ligaments, tendons, or joint capsules to bone, which is termed enthesitis, is a characteristic feature of spondyloarthropathy . Because of the relative inaccessibility of the enthesis, the inflammatory, microbiologic, and immunologic events at that site have been poorly defined . Recent magnetic resonance imaging studies have drawn attention to the ubiquitous nature of enthesitis in spondyloarthropathies, especially adjacent to synovial joints . This may have implications for the mechanisms of synovitis in spondyloarthropathies . Magnetic resonance imaging studies also suggest that enthesitis lesions may be extensive, which could explain the diffuse nature of bone changes seen in some patients with spondyloarthropathies . The importance of enthesitis as a skeletal phenomenon in spondyloarthropathies has gained further support from transgenic models in which either tumor necrosis factor-alpha or bone morphogenetic protein-6 overexpression result in entheseal-associated polyarthropathy. Clin Invest Med, 1999 Jun, 22(3), 111 - 8 Testing for HIV among patients with tuberculosis in Montreal; Geduld J et al.; OBJECTIVE: To determine the rate of HIV testing among patients with tuberculosis (TB) in Montreal, and to identify patient characteristics associated with physician screening patterns . Knowledge of local patterns of HIV testing among patients with TB could be used to facilitate the development of strategies to improve compliance with recommendations that all patients with TB be screened for HIV . DESIGN: Retrospective chart review . PATIENTS: All patients with TB reported to the Montreal Public Health Unit from 1992 to 1994 (ages 19 to 50) and from 1992 to 1995 (ages 18 and under) and for whom a chart could be reviewed . OUTCOME MEASURES: Patients with TB screened versus not screened for HIV infection, analyzed to determine which variables independently predict the likelihood of screening for HIV infection . RESULTS: Of the 376 patients with TB for whom data were available, 192 (51%) were screened for HIV . Of those, 33 (17%) had been tested before having received the diagnosis of TB . Multivariate analysis revealed that patients with TB who were male, aged 30 to 39, had a positive sputum smear, displayed at least 1 clinical symptom, received the TB diagnosis from a microbiologist or infectious disease specialist, or reported 1 or more HIV risk factors were more likely to be screened for HIV . CONCLUSIONS: HIV screening of patients with TB is selective, depending on both patient and physician characteristics . Physicians' awareness of recommendations needs to be increased to improve the diagnosis and treatment of TB/HIV co-infection. Rapid Commun Mass Spectrom, 1999 Jul, 13(13), 1334 - 1338 Determination of (15)N in (15)N-enriched nitrite and nitrate in aqueous samples by reaction continuous flow quadrupole mass spectrometry; Russow R; The (15)N tracer method is the most suitable method for studying complex N transformation processes in microbiology and biochemistry . It entails the constant determination of the (15)N abundance of the inorganic nitrogen (N) compounds nitrite and nitrate . However, (15)N analytical methods are time-consuming, difficult to automate, and require at least 10 microg of N per determination . An additional obstacle in the case of nitrite is that it usually only occurs in very small amounts (ppb) dwarfed by much larger quantities of nitrate (ppm) . More useful is an approach in which the N compound is selectively converted into a gaseous form suitable for direct measurement by mass spectrometry . By using this 'reaction continuous-flow mass spectrometry' (R/CFMS) we developed methods for the (15)N determination of nitrite and nitrate from tracer experiment samples, i.e . artificially enriched in (15)N . Because both methods are based on the same principle, one continuous flow setup connected directly to a quadrupole mass spectrometer for all determinations was used . Nitrite and nitrate are reduced to NO by iodide and titanium(III) chloride, respectively . The technique developed ensures a precision of relative standard deviation </=3% if at least 0.5 and 2 microg N with an abundance of >/=1 at.% are to be measured for nitrite and nitrate, respectively . Stat Med, 1999 Jul 15, 18(13), 1641 - 56 A test to detect replication in HIV serological data labelled by birth date based on the number of matching pairs in a sample; Greenhalgh D et al.; Diagnoses of HIV infection are reported to the Public Health Laboratory Service (PHLS) by microbiologists through a voluntary confidential surveillance scheme . Names are not recorded on the database but the date of birth of the individual concerned is usually available . This paper discusses a statistical method to detect repeated counting of individuals in these and similar data based on the number of matching pairs in the sample . The test is based on the theoretical result that the null hypothesis of all birth dates equally likely and all individuals distinct minimizes the expected number of matching pairs in the sample . Five of the 16 birth years in the sample taken in 1991 show evidence of more replication than would be expected by chance using a 5 per cent level test . When the test is repeated taking into account a small but statistically significant seasonal variation in the birth rate, the results are very similar . J Clin Microbiol, 1999 Aug, 37(8), 2587 - 91 Evaluation of the PrimeCapture CMV DNA detection plate system for detection of cytomegalovirus in clinical specimens; Davoli EH et al.; With the availability of anticytomegalovirus (CMV) therapeutic agents, rapid detection of CMV is important in the care and management of the immunosuppressed patient . The PrimeCapture CMV DNA Detection Plate System (PC-PCR) was evaluated for the detection of CMV in blood and cerebrospinal fluid (CSF) . The resolution of discordant results was performed by consensus testing utilizing a combination of conventional cell culture (TC-CPE), the CMV-antigenemia (CMV-Ag) assay, one or more in-house CMV nested PCR assays, and/or patient evaluation and follow-up . Of 51 blood specimens from 34 patients, 23 (45%) were identified as true positives . PC-PCR was significantly more sensitive than the CMV-Ag assay, TC-CPE, or a combination of both tests . The sensitivities, specificities, positive predictive values (PPV), and negative predictive values (NPV) for PC-PCR, the CMV-Ag assay, TC-CPE, and a combination of CMV-Ag and TC-CPE were 78, 75, 72, 81%; 46, 100, 100, 70%; 39, 100, 100, 67%; and 58, 100, 100, 73%, respectively . CMV was not detected or isolated in CSF, resulting in a combined PC-PCR sensitivity, specificity, PPV, and NPV of 77, 90, 68, and 93%, respectively . Among those laboratorians considering the incorporation of molecular CMV diagnostics into their clinical microbiology or virology laboratories, the CMV PC-PCR offers a relatively simple-to-perform and sensitive assay system. Diagn Microbiol Infect Dis, 1999 Jul, 34(3), 245 - 62 Cryptosporidia and microsporidia--waterborne diseases in the immunocompromised host; Franzen C et al.; Cryptosporidia and microsporidia are emerging parasitic pathogens in immunocompetent and immunocompromised individuals . Cryptosporidium infects several wild and domestic animals that excrete oocysts into the environment and contaminated water represents the major source of infection for humans . Waterborne transmission of Cryptosporidium is a major risk for humans and appropriate measures have to be taken to protect immunocompetent and immunocompromised individuals to become infected . For microsporidia, the sources and ways of transmission are not well documented . Although several animal hosts have been identified recently, the relevant reservoirs of human microsporidia are still unknown . Also, the routes of spreading are unknown . Is microsporidiosis a zoonotic disease that will be transmitted through close contact with infected animals or is contaminated surface water responsible for transmission and represents a relevant reservoir? This review is designed to give information on these two emerging intestinal parasites in a format that will be useful to clinical microbiologists, physicians interested in infectious diseases, and public health personnel. Commun Dis Public Health, 1999 Jun, 2(2), 151 - 2 Roles and responsibilities of safety officers and advisers; Tearle P; Health and safety should be integrated into all practices and procedures at work . This article looks at the differences between safety officers/advisers, also known as safety practitioners and appointed safely representatives, examining their roles and responsibilities within working environments in general and specifically in microbiology laboratories and offices. Commun Dis Public Health, 1999 Jun, 2(2), 149 - 50 Future accreditation of clinical microbiology laboratories; Djemal K; Accreditation organisations in the United Kingdom (UK) are seeking to harmonise their practices to comply with internationally recognized standards . As organisations and laboratories work towards achieving alignment with the international accreditation standards, the standards currently applied in clinical microbiology laboratories are likely to change . The enforcement of international standards may demand extra resources to accommodate the necessary changes and it is important that laboratory staff become aware of the potential influence of international accreditation standards on clinical microbiology. Commun Dis Public Health, 1999 Jun, 2(2), 122 - 5 Enterovirus infections in England and Wales: laboratory surveillance data: 1975 to 1994; Maguire HC et al.; Microbiology laboratories in England and Wales reported 40,366 culture confirmed isolates of echovirus (24,628; 61%) and coxsackievirus (B 11,714; 29%, A 4024; 10%) infections to the PHLS Communicable Disease Surveillance Centre (CDSC) in the 20 years from 1975 to 1994 . Nearly half of the organisms were isolated from faeces, and 5741 were isolated from cerebrospinal fluid (75% of them echovirus, 13% coxsackie B, and 12% coxsackie A) . Isolation rates for all enteroviruses were highest among infants aged 1 to 2 months . Sixty per cent of patients were aged under 5 years, 10% 5 to 9 years, and only 6% 35 years or over . Predominant serotypes were similar to those reported in other countries including the United States, Finland, and Belgium . Seventy-one per cent of reports were made between July and mid December . Periodicity varied between groups and serotypes: some demonstrated peaks at intervals of two to five years . There was evidence of spread of epidemic serotypes across Europe in certain years . Data collected between March and May each year enabled the strains circulating in the following 'season' to be predicted . Such information might be used to warn clinicians to anticipate particular clinical presentations. Circulation, 1999 Jul 13, 100(2), 155 - 63 Subunit expression of the cardiac L-type calcium channel is differentially regulated in diastolic heart failure of the cardiac allograft; Hullin R et al.; BACKGROUND: Left ventricular diastolic dysfunction is a major cause of cardiac allograft failure . Multimeric L-type calcium channels (alpha1-, alpha2/delta-, and beta-subunits) are essential for excitation/contraction coupling in the heart . Their gene expression was studied in allografts that developed diastolic heart failure . METHODS AND RESULTS: mRNA levels of calcium channel subunits were measured by competitive reverse transcriptase-polymerase chain reaction in microbiopsy samples from the interventricular septum . Size and tissue variabilities between biopsy samples were assessed by determination of cardiac calsequestrin mRNA levels . In the cardiac allografts studied, mRNA levels in microbiopsy samples were considered to represent left ventricular gene expression, because septal and left ventricular gene expression in Northern blots was equivalent, and left ventricles contracted homogeneously . Biopsy samples (n=72) were taken from allografts with normal left ventricular end-diastolic pressure (LVEDP; 8 to 13 mm Hg; n=30), moderately elevated LVEDP (14 to 18 mm Hg; n=26), and elevated LVEDP (19 to 28 mm Hg; n=16) . Increased LVEDP was related to slowed diastolic relaxation determined by the time constant tau (r2=0.86), whereas systolic performance (dP/dt; ejection fraction) was preserved . With increasing LVEDP, mRNA levels of the pore-forming alpha1c-subunit (n=15) and of the regulatory alpha2/delta-subunit (n=17) remained unchanged but decreased exponentially (r2=-0.83) for the regulatory beta-subunit (n=40) . Compared with cardiac allografts with normal LVEDP (n=15), beta-subunit mRNA level was reduced by 75% at elevated LVEDP (n=9; P=0.012) . In an explanted, diastolically failing cardiac allograft, beta-subunit expression was reduced correspondingly by 72% and 76% on the mRNA level in septal and left ventricular myocardium and by 80% on the protein level . CONCLUSIONS: The downregulated expression of the calcium channel beta-subunit might contribute to altered calcium handling in diastolically failing cardiac allografts. Luminescence, 1999 Jan-Feb, 14(1), 19 - 22 Application of a new high sensitivity luminometer for industrial microbiology and molecular biology; Andreotti PE et al.; A compact new luminometer (FB12) has been developed based on a 370-630 nm photon counter and measuring chamber that can accommodate a range of sample formats . The FB12 permits measurements as low as 1000 molecules of luciferase in reporter gene assays . Its sensitivity for ATP is limited by reagent background . If ATP assay reagents had no chemical background, 2 fg of ATP could be detected using 3 SD of instrument background as the detection limit . The FB12 has a dynamic range of six decades and operates under its own microprocessor programme or protocol-based PC software that is integrated with Microsoft(R) Excel(R) . An injector port above the sample measuring position allows connection of external reagent injectors . Applications are performed using protocols provided with the FB12 or user defined protocols . Examples are presented that illustrate use of the instrument for research and industrial applications . Monaldi Arch Chest Dis, 1999 Apr, 54(2), 115 - 9 Bronchoalveolar lavage and transbronchial lung biopsy in alveolar and/or ground-glass opacification; Cazzato S et al.; In order to assess the diagnostic yield of bronchoalveolar lavage (BAL) and transbronchial lung biopsy (TBB) in pulmonary diseases with a ground-glass and/or alveolar pattern on high-resolution computed tomography (HRCT) scan, a prospective study was undertaken . Thirty-six patients (17 males, 19 females), mean age 53 yrs, selected on the basis of the presence of an alveolar and/or ground-glass pattern on chest HRCT scan, were submitted to fibreoptic bronchoscopy . All patients underwent BAL . TBBs were performed in 33 cases . A specific diagnosis was achieved, taking into account data obtained by means of serology, microbiology, cytology and histopathology in appropriate clinical settings . Twelve (33%) patients only had the appearance of a ground-glass opacity, whereas 24 (67%) had associated areas of airspace consolidation . BAL was performed in all cases and gave a definitive diagnosis in 21 (58%) . The diagnostic yield of BAL in patients with only ground-glass opacities was no different from that in those patients also showing areas of alveolar consolidation (58 versus 58%) . In eight patients (six with ground-glass opacity and two with alveolar consolidation), BAL provided useful but not definitive information . In these patients, a definitive diagnosis was achieved by means of TBB in seven cases and by open lung biopsy in one case . TBB was performed in 33 out of 36 patients and gave positive results in 25 (76%) . The diagnostic yield of TBB in patients showing areas of alveolar consolidation was significantly higher than in those with pure ground-glass opacity, i.e . 95% (21 of 22) and 36% (4 of 11) respectively (p < 0.001) . BAL and TBB were performed during the same bronchoscopy in 33 patients, and an accurate diagnosis was achieved in 30 (91%) . Overall, the diagnostic yield of TBB (76%) and BAL (56%) did not differ significantly in the whole patient group (p = 0.12), or in patients with a ground-glass opacification (58 versus 36%, p = 0.3) . However, in patients with areas of alveolar consolidation, the diagnostic sensitivity of TBB (95%) was significantly greater than the diagnostic sensitivity of BAL (54%) (p = 0.03) . In conclusion this study shows that high-resolution computed tomography can be helpful in predicting the diagnostic accuracy of bronchological procedures, in particular of bronchoalveolar lavage and transbronchial biopsy, and that alveolar and/or ground-glass are favourable patterns for these diagnostic tools. Am J Respir Crit Care Med, 1999 Jul, 160(1), 346 - 8 Initial microbiologic studies did not affect outcome in adults hospitalized with community-acquired pneumonia; Sanyal S et al.; Microbiologic studies (MBSs) fail to identify a specific pathogen in more than 50% of patients with community-acquired pneumonia (CAP) . The 1993 American Thoracic Society guideline (ATS-GL) for the management of CAP advised selecting initial antibiotic regimens based on severity of illness and comorbidities . Our study evaluated the role of initial MBS in adult patients hospitalized with CAP and treated according to the ATS-GL . In 184 patients hospitalized at our facility for CAP in 1996, and treated according to the ATS-GL, 25 (14%) failed to respond to initial antibiotic regimens . In these nonresponders, there was no difference in mortality between those in whom antibiotics were changed empirically, and those with MBS-guided changes . We conclude that initial MBS may not be warranted in many adult patients admitted for CAP . Exceptions include patients with conditions that predispose to less common, more resistant pathogens. J Appl Microbiol, 1999 Jun, 86(6), 999 - 1007 Low level chemiluminescence from liquid culture media; Vogel R et al.; Low level chemiluminescence (CL) can be observed from autoclaved liquid culture media, as used in microbiology . The light emission is oxygen-dependent and arises from reactions following auto-oxidation of reducing Maillard products which are formed during autoclaving . The inhibition of this CL by radical scavengers and antioxidants has been studied . As superoxide radicals and hydrogen peroxide are predominantly involved in the initiation of the CL, the investigation of CL from culture media offers a convenient tool for the detection of exogenous (medium-mediated) oxidative stress being imposed onto micro-organisms in culture . Transition metal ions showed, dependent on concentration, both inhibitory and stimulating effects on the CL, which was also affected by the presence of complexing agents . Iron porphyrins and related complexes displayed a very efficient quenching of the CL, which may be of particular importance, as aerobic micro-organisms have been previously shown to be very efficient in quenching the CL from culture media. Appl Environ Microbiol, 1999 Jul, 65(7), 3084 - 94 Photosynthetic bradyrhizobia from Aeschynomene spp . are specific to stem-nodulated species and form a separate 16S ribosomal DNA restriction fragment length polymorphism group; Molouba F et al.; We obtained nine bacterial isolates from root or collar nodules of the non-stem-nodulated Aeschynomene species A . elaphroxylon, A . uniflora, or A . schimperi and 69 root or stem nodule isolates from the stem-nodulated Aeschynomene species A . afraspera, A . ciliata, A . indica, A . nilotica, A . sensitiva, and A . tambacoundensis from various places in Senegal . These isolates, together with 45 previous isolates from various Aeschynomene species, were studied for host-specific nodulation within the genus Aeschynomene, also revisiting cross-inoculation groups described previously by D . Alazard (Appl . Environ . Microbiol . 50:732-734, 1985) . The whole collection of Aeschynomene nodule isolates was screened for synthesis of photosynthetic pigments by spectrometry, high-pressure liquid chromatography, and thin-layer chromatography analyses . The presence of puf genes in photosynthetic Aeschynomene isolates was evidenced both by Southern hybridization with a Rhodobacter capsulatus photosynthetic gene probe and by DNA amplification with primers defined from photosynthetic genes . In addition, amplified 16S ribosomal DNA restriction analysis was performed on 45 Aeschynomene isolates, including strain BTAi1, and 19 reference strains from Bradyrhizobium japonicum, Bradyrhizobium elkanii, and other Bradyrhizobium sp . strains of uncertain taxonomic positions . The 16S rRNA gene sequence of the photosynthetic strain ORS278 (LMG 12187) was determined and compared to sequences from databases . Our main conclusion is that photosynthetic Aeschynomene nodule isolates share the ability to nodulate particular stem-nodulated species and form a separate subbranch on the Bradyrhizobium rRNA lineage, distinct from B . japonicum and B . elkanii. Appl Environ Microbiol, 1999 Jul, 65(7), 2987 - 93 Mechanism of pyrite dissolution in the presence of Thiobacillus ferrooxidans; Fowler TA et al.; In spite of the environmental and commercial interests in the bacterial leaching of pyrite, two central questions have not been answered after more than 35 years of research: does Thiobacillus ferrooxidans enhance the rate of leaching above that achieved by ferric sulfate solutions under the same conditions, and if so, how do the bacteria affect such an enhancement? Experimental conditions of previous studies were such that the concentrations of ferric and ferrous ions changed substantially throughout the course of the experiments . This has made it difficult to interpret the data obtained from these previous works . The aim of this work was to answer these two questions by employing an experimental apparatus designed to maintain the concentrations in solution at a constant value . This was achieved by using the constant redox potential apparatus described previously (P . I . Harvey, and F . K . Crundwell, Appl . Environ . Microbiol . 63:2586-2592, 1997; T . A . Fowler, and F . K . Crundwell, Appl . Environ . Microbiol . 64:3570-3575, 1998) . Experiments were conducted in both the presence and absence of T . ferrooxidans, maintaining the same conditions in solution . The rate of dissolution of pyrite with bacteria was higher than that without bacteria at the same concentrations of ferrous and ferric ions in solution . Analysis of the dependence of the rate of leaching on the concentration of ferric ions and on the pH, together with results obtained from electrochemical measurements, provided clear evidence that the higher rate of leaching with bacteria is due to the bacteria increasing the pH at the surface of the pyrite. Curr Opin Microbiol, 1999 Jun, 2(3), 299 - 305 Ribosomal DNA sequencing as a tool for identification of bacterial pathogens; Kolbert CP et al.; Conventional methods for the identification and characterization of clinical isolates of bacterial pathogens sometimes fall short when such isolates exhibit unusual phenotypic profiles . Recent advances in DNA sequencing technology have greatly enhanced the ability of the microbiologist to determine the identity of a bacterial isolate . Given the relative objectivity of DNA sequence information and growing availability of sequence information databases, a significant movement is now afoot to use molecular methods for the identification of clinical pathogens. Cardiovasc Intervent Radiol, 1999 May-Jun, 22(3), 246 - 8 Post-traumatic pseudocyst of the spleen: sclerotherapy with ethanol; Volk M et al.; We report a case of successful percutaneous treatment of a chronic post-traumatic splenic pseudocyst using alcohol as the sclerosing agent . A 26-year-old man presented with a symptomatic cystic mass located in the spleen . Aspiration of 300 ml of fluid was only temporarily effective, and therefore a drainage catheter was placed 3 days later . After histopathologic and microbiologic exclusion of a malignant or infectious origin, local sclerotherapy with alcohol was performed because of recurrence after percutaneous drainage . This therapy was repeated six times within 2 weeks . Two weeks later, the remaining volume was determined to be 16 ml . Six months after treatment the cyst was no longer visible . To our knowledge this is the first case of a chronic post-traumatic splenic cyst treated with alcohol . Percutaneous sclerotherapy of a symptomatic post-traumatic splenic pseudocyst may be an alternative to surgical treatment. Retina, 1999, 19(3), 213 - 7 Hypodermic needles: a new source of penetrating ocular trauma in Indian children; Jalali S et al.; PURPOSE: To study the clinico-microbiologic profile and visual prognosis of ocular injuries caused by disposable hypodermic needles used by children to squirt water . METHODS: We analyzed 19 consecutive cases of hypodermic needle injury seen at our institute . RESULTS: The average age of the patients was 10.3 years (range, 4-20 years) . A small self-sealed corneal or scleral laceration was seen in 11 eyes; in 8 eyes, the site of injury was occult . Initial visual acuity was no light perception (3 eyes) or hand motion or light perception (16 eyes) . Surgery in 18/19 eyes included vitrectomy with intraocular antibiotic injections for endophthalmitis (14 eyes), evisceration for panophthalmitis (2 eyes), and cataract extraction for traumatic cataract (2 eyes) . Final visual acuity was no light perception or light perception only in 10 eyes, 20/400-20/60 in three eyes, and 20/40 or better in six eyes . CONCLUSIONS: Severe ocular morbidity may result from improper disposal of hypodermic needles. Cell Mol Life Sci, 1999 May, 55(5), 751 - 70 Immune responses to DNA vaccines; Kowalczyk DW et al.; DNA vaccines, based on plasmid vectors expressing an antigen under the control of a strong promoter, have been shown to induce protective immune responses to a number of pathogens, including viruses, bacteria and parasites . They have also displayed efficacy in treatment or prevention of cancer, allergic diseases and autoimmunity . Immunologically, DNA vaccines induce a full spectrum of immune responses that include cytolytic T cells, T helper cells and antibodies . The immune response to DNA vaccines can be enhanced by genetic engineering of the antigen to facilitate its presentation to B and T cells . Furthermore, the immune response can be modulated by genetic adjuvants in the form of vectors expressing biologically active determinants or by more traditional adjuvants that facilitate uptake of DNA into cells . The ease of genetic manipulation of DNA vaccines invites their use not only as vaccines but also as research tools for immunologists and microbiologists. Artif Organs, 1999 May, 23(5), 460 - 2 Experimental wound healing with electrical stimulation; Reger SI et al.; The effect of alternating current (AC) and direct current (DC) stimulation was studied on experimental pressure ulcer healing in a new monoplegic pig model . The study was conducted in 30 healthy young Hanford minipigs . The rate of wound healing, histology, vascularization, collagen formation, microbiology, perfusion, and the mechanical strength of the healed wounds were studied . Normal pigskin was compared to denervated control and denervated AC and DC stimulated healed skin . Hind limb denervation was by right unilateral extradural rhizotomies from the L2 to S1 nerve roots . Reproducible uniformly controlled Stage III or higher tissue ulcers were created . When compared to the control wounds, both the AC and DC stimulated wounds showed reduced healing time and increased perfusion in the early phases of healing . DC stimulation reduced the wound area more rapidly than AC, but AC stimulation reduced the wound volume more rapidly than DC . The electrical stimulation did not reduce the strength of the healing wounds below those of the nonstimulated controls . The applied current appears to orient new collagen formation even in the absence of neural influences. Przegl Lek, 1999, 56(1), 72 - 5 {The effectiveness of conservative treatment of cervical lesions using the LLETZ and CO2 laser}; Lisowski P et al.; Development made on the carcinogenesis process of the cervical lesions and increased detection of the early precancerous lesions enable discontinuance of radical treatments for non-radical techniques which it is of vital importance to young women of the child-bearing capacity . The aim of this study was to determine the efficacy of the non-radical treatment of the cervical lesions using LLETZ procedure (Large Loop Excision of Transition Zone) and laser CO2 vaporisation . 2046 women aged 18-46 who were diagnosed for cervical lesions were treated in the Institute of Obstetrics and Gynaecology Medical School of Bialystok in the years 1994-97 . 216 of which were histologically confirmed for CIN I-III diagnosis . The choice between LLETZ or laser CO2 was made based on a pre-treatment examination (cytology, colposcopy, microbiology test and punch biopsy) . The final results were evaluated from 6 months to 4 years after the treatment . The effectiveness of CO2 laser was 94.6% and was similar to LLETZ--96.4% . In spite of almost complete agreement in both procedures, the LLETZ seems to be more preferred because of the possibility of histological post-treatment verification. Med Clin (Barc), 1999 May 15, 112(17), 641 - 5 {The usefulness of the bone marrow examination in the etiological diagnosis of prolonged fever in patients with HIV infection}; Fernandez-Aviles F et al.; BACKGROUND: To analyze the usefulness of bone marrow (BM) study in the diagnosis of fever of unknown origin (FUO) in patients infected by the human immunodeficiency virus (HIV) in a single center during a period of 42 months . PATIENTS AND METHODS: 182 episodes of FUO in 162 patients p3th HIV infection were studied . Age, sex, risk factor for HIV infection, hemoglobin level, counts of leucocytes, neutrophils, lymphocytes, CD4 positive lymphocytes, platelets and levels of hepatic enzymes, albumin and beta 2-microglobulin were studied . BM aspirate was performed in all episodes for cytologic and microbiologic examination, and BM biopsy was done in 43 . Analysis of factors related with the probability of diagnosis by BM examination was carried out . RESULTS: The median age was 33 years (range, 22-70), and 123 were males . Drug abuse was the most frequent risk factor for HIV infection (63%) . One hundred thirty patients had previous AIDS diagnosis before they were evaluated for unexplained fever . A specific diagnosis was achieved in 161 episodes (88%) and the most frequent diagnoses were Mycobacterium spp . (55%) and Leishmania spp . (14%) infections . Fifty-four episodes (30%) were diagnosed by BM examination, and in 36 (20%) BM study was the only diagnostic tool . Examination of the BM aspirate yielded the diagnosis in 40 out of the 178 episodes (13%), whereas BM biopsy was a diagnostic tool in 8 (19%); in 9 additional cases (21%) granulomas were observed . Microbiologic study of BM smears for mycobacterial infections was positive in 28 of the 143 episodes (19%), and the culture for Leishmania was positive in 2 out of the 42 cases . The presence of thrombocytopenia (< 75 x 10(9)/l) and elevated serum levels of aspartate-aminotransferase (AST) (> 100 U/l) were the factors associated with a high probability to obtain the diagnosis through BM study . CONCLUSIONS: In patients infected by the HIV and unexplained fever, BM examination is an useful procedure for the diagnosis, particularly in areas where infections by Mycobacterium spp . and Leishmania are prevalent . So that, in our setting, systematic use of this procedure is justified for diagnosis of FUO in those patients. Arch Dis Child, 1999 Jul, 81(1), 64 - 7 Vulvovaginitis: clinical features, aetiology, and microbiology of the genital tract; Jaquiery A et al.; AIM: To clarify the contribution of clinical and environmental factors and infection to the aetiology of vulvovaginitis in premenarchal girls, and to determine clinical indicators of an infectious cause . DESIGN: It was necessary first to define normal vaginal flora . Cases were 50 premenarchal girls > 2 years old with symptoms of vulvovaginitis; 50 controls were recruited from girls in the same age group undergoing minor or elective surgery . RESULTS: Interview questionnaire showed no difference between cases and controls in regards to hygiene practices, exposure to specific irritants, or history of possible sexual abuse . Normal vaginal flora was similar to that described in previous studies, with the exception of organisms likely to be associated with sexual activity . 80% of cases had no evidence of an infectious cause . In the 10 cases in whom an infectious cause was found, there was significantly more visible discharge and distinct redness of the genital area on examination compared with other cases . CONCLUSIONS: The findings suggest that vulvovaginitis in this age group is not usually infectious or necessarily related to poor hygiene, specific irritants or sexual abuse, although any of these can present with genital irritation . The possibility of sexual abuse should always be considered when a child presents with genital symptoms, but our data indicate it is not a common contributing factor . Infection is generally associated with vaginal discharge and moderate or severe inflammation. J Zoo Wildl Med, 1999 Mar, 30(1), 100 - 10 Brucella-induced abortions and infection in bottlenose dolphins (Tursiops truncatus); Miller WG et al.; Two bottlenose dolphins (Tursiops truncatus) aborted fetuses that died as a result of Brucella infection . Brucella placentitis occurred in both cases . Infected placenta and vaginal/uterine fluids may transmit Brucella species to other cetaceans . In a third case, an identical organism was cultured from lung necropsy tissue of an adult female T . truncatus . Microbiology, specific polymerase chain reaction, and pulsed-field gel electrophoresis results supported the designation of an additional genomic group(s), Brucella delphini, for isolates adapted to T . truncatus . Current serologic diagnostic tests reliable for known Brucella species are unreliable in detecting dolphin brucellosis . Our findings, together with previous reports, suggest that dolphin brucellosis is a naturally occurring disease that can adversely impact reproduction in cetaceans . The zoonotic significance of cetacean brucellosis is unknown, although the disease has not been reported in people who have frequent contact with dolphins . Further studies on the zoonotic aspects, distribution, prevalence, virulence, and impact of this disease in cetaceans and other marine mammal species are needed. J Clin Microbiol, 1999 Jul, 37(7), 2241 - 7 Validity of interpretation criteria for standardized Western blots (immunoblots) for serodiagnosis of Lyme borreliosis based on sera collected throughout Europe; Hauser U et al.; Western blotting (WB; immunoblotting) is a widely used tool for the serodiagnosis of Lyme borreliosis (LB), but so far, no generally accepted criteria for performance and interpretation have been established in Europe . The current study was preceeded by a detailed analysis of WB with whole-cell lysates of three species of Borrelia burgdorferi sensu lato (U . Hauser, G . Lehnert, R . Lobentanzer, and B . Wilske, J . Clin . Microbiol . 35:1433-1444, 1997) . In that study, interpretation criteria for a positive WB result were developed with the data for 330 serum samples (from patients with LB in different stages {n = 189} and from a control group {n = 141}) originating mostly from southern Germany . In the present work, the interpretation criteria for strains PKo (Borrelia afzelii) and PBi (Borrelia garinii) developed in the previous study were reevaluated with 224 serum samples (from patients with LB in different stages {n = 97} and from a control group {n = 127}) originating from throughout Europe that were provided by the European Union Concerted Action on Lyme Borreliosis (EUCALB) . De novo criteria were developed on the basis of the reactivities of the EUCALB sera and were evaluated with the data for the samples from southern Germany . Comparison of all results led to the following recommendations: For WB for immunoglobulin G (IgG), at least two bands among p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo and at least one band among p83/100, p39, p30, OspC, p21, and p17b for PBi; for WB for IgM, at least one band among p39, OspC, and p17 or a strong p41 band for PKo and at least one band among p39 and OspC or a strong p41 band for PBi . WB with PKo was the most sensitive, and this strain is recommended for use in WB for the serodiagnosis of LB throughout Europe. J Hosp Infect, 1999 May, 42(1), 1 - 6 Laboratory role in the management of hospital acquired infections; Wilson MP et al.; The microbiology laboratory has many important roles . It must collaborate with the infection control team on the investigations of outbreaks . During outbreaks, it must save relevant samples, look for reservoirs and undertake typing techniques, all of which should be timely . New technology should be available to detect, identify and characterize micro-organisms . Molecular biological techniques have enhanced the speed and sensitivity of detection methods and have allowed the laboratory to identify organisms that do not grow or grow slowly in culture . Molecular techniques also enable the microbiologist to identify antibiotic resistance genes and to 'fingerprint' hospital organisms, thereby facilitating studies of nosocomial transmission. Adolesc Med, 1995 Feb, 6(1), 45 - 54 Infectious Hepatitis A and Enteric Hepatitis E; Rose E et al.; Infectious hepatitis A and enteric hepatitis E have a peak incidence in late adolescence or young adulthood . Using a 14-year-old boy as an example, the authors trace the course of hepatitis A through the preicteric and icteric phases, the development of persistent jaundice, relapse, and liver failure . The discussion of hepatitis E addresses microbiology and clinical course of the condition. Microbiol Mol Biol Rev . 1999 Jun;63(2):265. A paean to Microbiology and Molecular Biology Reviews; Schaechter M; This article celebrates the accomplishments of Microbiology and Molecular Biology Reviews from its early days to the present time . The importance of this journal in the professional lives of microbiologists is emphasized, and examples of outstanding reviews are presented. Int J Food Microbiol, 1999 Mar 1, 47(1-2), 141 - 6 Predicting fungal growth: the effect of water activity on Penicillium roqueforti; Valik L et al.; The effect of water activity on the colony growth of Penicillium roqueforti is studied by predictive modelling techniques . Measured colony diameter growth curves are fitted to estimate the growth rate and lag phase of the curves . The colony growth rate was modelled by a quadratic function of transformed water activity (a(w)) values, as suggested by Baranyi et al . (Food Microbiol . 10 (1993) 43-59) . The lag time was modelled as a function of water activity, by means of the sum of a constant term and a hyperboloid function of a(w) raised to the second power . The lag-phase of Penicillium roqueforti was found insensitive to the water activity in the range of its higher (a(w) > 0.92) values. Clin Lab Sci, 1999 Jan-Feb, 12(1), 12 - 3 The use of interactive technology in the classroom; Kresic P; This article discusses the benefits that clinical laboratory science students and instructors experienced through the use of and integration of computer technology, microscopes, and digitizing cameras . Patient specimens were obtained from the participating clinical affiliates, slides stained or wet mounts prepared, images viewed under the microscope, digitized, and after labeling, stored into an appropriate folder . The individual folders were labeled as Hematology, Microbiology, Chemistry, or Urinalysis . Students, after obtaining the necessary specimens and pertinent data, created case study presentations for class discussions . After two semesters of utilizing videomicroscopy/computer technology in the classroom, students and instructors realized the potential associated with the technology, namely, the vast increase in the amount of organized visual and scientific information accessible and the availability of collaborative and interactive learning to complement individualized instruction . The instructors, on the other hand, were able to provide a wider variety of visual information on individual bases . In conclusion, the appropriate use of technology can enhance students' learning and participation . Increased student involvement through the use of videomicroscopy and computer technology heightened their sense of pride and ownership in providing suitable information in case study presentations . Also, visualization provides students and educators with alternative methods of teaching/learning and increased retention of information. Adolesc Med, 1990 Oct, 1(3), 565 - 582 Sexually Transmitted Diseases in Homosexual Youth; Remafedi G; The prevalence of sexually transmitted diseases (STDs) in homosexual populations is due to a complex interrelationship of host and pathogen factors . This chapter explores the risk factors, epidemiology, microbiology, clinical aspects, and prevention of STDs in homosexual youth . This discussion focuses on male adolescents, because STDs are uncommon among exclusively homosexual women. Appl Environ Microbiol, 1999 Jun, 65(6), 2341 - 9 Effect of temperature on structure and function of the methanogenic archaeal community in an anoxic rice field soil; Chin KJ et al.; Soil temperatures in Italian rice fields typically range between about 15 and 30 degrees C . A change in the incubation temperature of anoxic methanogenic soil slurry from 30 degrees C to 15 degrees C typically resulted in a decrease in the CH4 production rate, a decrease in the steady-state H2 partial pressure, and a transient accumulation of acetate . Previous experiments have shown that these changes were due to an alteration of the carbon and electron flow in the methanogenic degradation pathway of organic matter caused by the temperature shift (K . J . Chin and R . Conrad, FEMS Microbiol . Ecol . 18:85-102, 1995) . To investigate how temperature affects the structure of the methanogenic archaeal community, total DNA was extracted from soil slurries incubated at 30 and 15 degrees C . The archaeal small-subunit (SSU) rRNA-encoding genes (rDNA) of these environmental DNA samples were amplified by PCR with an archaeal-specific primer system and used for the generation of clone libraries . Representative rDNA clones (n = 90) were characterized by terminal restriction fragment length polymorphism (T-RFLP) and sequence analysis . T-RFLP analysis produced for the clones terminally labeled fragments with a characteristic length of mostly 185, 284, or 392 bp . Sequence analysis allowed determination of the phylogenetic affiliation of the individual clones with their characteristic T-RFLP fragment lengths and showed that the archaeal community of the anoxic rice soil slurry was dominated by members of the families Methanosarcinaceae (185 bp) and Methanosaetaceae (284 bp), the kingdom Crenarchaeota (185 or 284 bp), and a novel, deeply branching lineage of the (probably methanogenic) kingdom Euryarchaeota (392 bp) that has recently been detected on rice roots (R . Grosskopf, S . Stubner, and W . Liesack, Appl . Environ . Microbiol . 64:4983-4989, 1998) . The structure of the archaeal community changed when the temperature was shifted from 30 degrees C to 15 degrees C . Before the temperature shift, the clones (n = 30) retrieved from the community were dominated by Crenarchaeota (70%), "novel Euryarchaeota" (23%), and Methanosarcinacaeae (7%) . Further incubation at 30 degrees C (n = 30 clones) resulted in a relative increase in members of the Methanosarcinaceae (77%), whereas further incubation at 15 degrees C (n = 30 clones) resulted in a much more diverse community consisting of 33% Methanosarcinaceae, 23% Crenarchaeota, 20% Methanosaetaceae, and 17% novel Euryarchaeota . The appearance of Methanosaetaceae at 15 degrees C was conspicuous . These results demonstrate that the structure of the archaeal community in anoxic rice field soil changed with time and incubation temperature. J Clin Pathol, 1999 Jan, 52(1), 54 - 60 Use of bar code readers and programmable keypads to improve the speed and accuracy of manual data entry in the clinical microbiology laboratory: experience of two laboratories; Shaw R et al.; AIM: To assess the effect of the use of bar code readers and programmable keypads for entry of specimen details and results in two microbiology laboratories . METHODS: The solutions selected in each laboratory are described . The benefits resulting from the implementation were measured in two ways . The speed of data entry and error reduction were measured by observation . A questionnaire was completed by users of bar codes . RESULTS: There were savings in time and in reduced data entry errors . Average time to enter a report by keyboard was 21.1 s v 14.1 s for bar coded results entry . There were no observed errors with the bar code readers but 55 errors with keystroke entries . The laboratory staff of all grades found the system fast, easy to use, and less stressful than conventional keyboard entry . CONCLUSIONS: Indirect time savings should accrue from the observed reduction in incorrectly entered data . Any microbiology laboratory seeking to improve the accuracy and efficiency of data entry into their laboratory information systems should consider the adoption of this technology which can be readily interfaced to existing terminals. Cornea, 1999 May, 18(3), 257 - 61 Single culture media in infectious keratitis; Waxman E et al.; PURPOSE: To evaluate the usefulness of the various culture media used in the traditional workup in infectious keratitis . METHODS: Microbiology data sheets from all corneal cultures performed at the University of California Davis Medical Center over a 1-year period were reviewed retrospectively . RESULTS: Bacterial cultures were sent in 76 cases . In 19 cases, culture specimens from ulcers were plated onto blood, chocolate, and inhibitory mold agar and were inoculated into an anaerobic medium . In 58 cases, blood and chocolate agar were sent . In 70% of cases, blood and chocolate agar provided identical information . Inhibitory mold agar was positive twice in 39 plates sent . A fungal pathogen had been identified on chocolate agar plates sent for these cases . CONCLUSION: In the evaluation of infectious keratitis, plating onto chocolate agar or blood agar alone is a reasonable alternative to sending multiple cultures. Br Dent J, 1999 Apr 10, 186(7), 345 - 7 Barriers to the use of a diagnostic oral microbiology laboratory by general dental practitioners; Roy KM et al.; OBJECTIVE: To identify barriers on the use of diagnostic microbiology facilities in general dental practice . DESIGN: A cross-sectional survey using a postal questionnaire . SETTING: Primary/secondary care interface between the diagnostic oral microbiology laboratory, University of Glasgow Dental Hospital and School, Glasgow and dental practitioners within the surrounding health boards, 1998 . SUBJECTS: All GDPs (797) within Argyll and Clyde, Ayrshire and Arran, Lanarkshire and Greater Glasgow Health Boards . MAIN OUTCOME MEASURES: The responses were expressed as both absolute and relative frequencies . RESULTS: Responses were received from 430 (55%) . The most frequent reason for failure to use the service was lack of information, with more than half of the respondents claiming to be unaware of the facility . Lack of request forms and sampling equipment were also viewed as barriers to using the service . CONCLUSIONS: The laboratory is failing to successfully communicate its role in addressing the growing burden of antibiotic resistance in the community and must be more proactive in encouraging appropriate use and increasing accessibility of the service to GDPs. Front Biosci, 1999 May 15, 4, e34 - 41 Inhibitor resistant class A beta-lactamases; Bonomo RA et al.; Beta-lactamase inhibitors (clavulanic acid, tazobactam, and sulbactam) greatly enhance the therapeutic efficacy of their partner antibiotics (amoxacillin, ampicillin, piperacillin, and ticarcillin) against common enteric and non-enteric organisms possessing class A beta-lactamases . Unfortunately, the number of class A enzymes being discovered that are resistant to these combinations is increasingly rapidly . The TEM and SHV class A beta-lactamases resistant to inhibitors have point mutations in critical amino acids important for catalysis . Compared to the wild type beta-lactamase, inhibitor resistant enzymes are inefficient at hydrolyzing benzylpenicillin, aminopenicillins, and cephalosporins . Nevertheless, hyper-production of these enzymes resulting from mutations in the promoter region can confer substantial levels of resistance . Understanding the microbiologic and kinetic properties of these inhibitor resistant class A beta-lactamases can lead to the design of more potent beta-lactam compounds as well as more effective inhibitors. Mikrobiol Z, 1999 Jan-Feb, 61(1), 3 - 31 {The patterns in the molecular biology of the adenoviruses (the results of a 20-year study at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine)}; Diachenko NS; Scientific data of the highest importance and priority concerning regularities of structural and functional organization of proteins of adenoviruses capsids and peculiarities of expression of the virus genome are as follows: New antigen determinants of hexon and adenovirus fiber have been discovered, their different nature (conformational or linear) and different orientation, depending on the spatial organization of proteins, have been proved; localization of some epitopes has been determined with the help of synthesised antigen-active peptides, imitating them . Some regularities of structural and functional organization of adenovirus hexon have been determined on the basis of comparative analysis of antigenic specificity and primary structure of proteins being apart in taxonomic respect . The conception of immunoactivation (infectivity neutralization) of adenoviruses has been developed, and a mathematical model of this process has been first proposed, which determines the impact of antibodies to several antigenic determinants of hexon and fiber as well as interferon and complement . The unknown peculiarities of the adenovirus genome expression were studied in the dynamics of productive infection or under the effect of modified nucleosides, proteolysis inhibitors and those of different nature promising for chemotherapy of adenovirus infection . Lymphotropicity of adenoviruses was established and a model of the mixed infection of lymphocytes with adenoviruses, HIV, and Epstein-Barr virus of the herpes virus family was proposed for the first time . It was determined that the mutual interference of viruses was developed at the process of a single or successive infection and this was important to understand AIDS immunopathogenesis . Data presented substantiate the ways of creation of modern efficient preparations for diagnosis, prophylaxis and chemotherapy of adenovirus infection. Arch Intern Med, 1999 May 10, 159(9), 970 - 80 Processes and outcomes of care for patients with community-acquired pneumonia: results from the Pneumonia Patient Outcomes Research Team (PORT) cohort study; Fine MJ et al.; BACKGROUND: Although understanding the processes of care and medical outcomes for patients with community-acquired pneumonia is instrumental to improving the quality and cost-effectiveness of care for this illness, limited information is available on how physicians manage patients with this illness or on medical outcomes other than short-term mortality . OBJECTIVES: To describe the processes of care and to assess a broad range of medical outcomes for ambulatory and hospitalized patients with community-acquired pneumonia . METHODS: This prospective, observational study was conducted at 4 hospitals and 1 health maintenance organization in Pittsburgh, Pa, Boston, Mass, and Halifax, Nova Scotia . Data were collected via patient interviews and reviews of medical records for 944 outpatients and 1343 inpatients with clinical and radiographic evidence of community-acquired pneumonia . Processes of care and medical outcomes were assessed 30 days after presentation . RESULTS: Only 29.7% of outpatients had 1 or more microbiologic tests performed, and only 5.7% had an assigned microbiologic cause . Although 95.7% of inpatients had 1 or more microbiologic tests performed, a cause was established in only 29.6% . Six outpatients (0.6%) died, and 3 of these deaths were pneumonia related . Of surviving outpatients, 8.0% had 1 or more medical complications . At 30 days, 88.9% (nonemployed) to 95.6% (employed) of the surviving outpatients had returned to usual activities, yet 76.0% of outpatients had 1 or more persisting pneumonia-related symptoms . Overall, 107 inpatients (8.0%) died, and 81 of these deaths were pneumonia related . Most surviving inpatients (69.0%) had 1 or more medical complications . At 30 days, 57.3% (non-employed) to 82.0% (employed) of surviving inpatients had returned to usual activities, and 86.1% had 1 or more persisting pneumonia-related symptoms . CONCLUSIONS: In this study, conducted primarily at hospital sites with affiliated medical education training programs, virtually all outpatients and most inpatients had pneumonia of unknown cause . Although outpatients had an excellent prognosis, pneumonia-related symptoms often persisted at 30 days . Inpatients had substantial mortality, morbidity, and pneumonia-related symptoms at 30 days. J Clin Microbiol, 1999 Jun, 37(6), 2106 - 8 Mycobacterium bovis BCG causing vertebral osteomyelitis (Pott's disease) following intravesical BCG therapy; Aljada IS et al.; We report a case of Mycobacterium bovis BCG vertebral osteomyelitis in a 79-year-old man 2.5 years after intravesical BCG therapy for bladder cancer . The recovered isolate resembled M . tuberculosis biochemically, but resistance to pyrazinamide (PZA) rendered that diagnosis suspect . High-pressure liquid chromatographic studies confirmed the diagnosis of M . bovis BCG infection . The patient was originally started on a four-drug antituberculous regimen of isoniazid, rifampin, ethambutol, and PZA . When susceptibility studies were reported, the regimen was changed to isoniazid and rifampin for 12 months . Subsequently, the patient was transferred to a skilled nursing facility for 3 months, where he underwent intensive physical therapy . Although extravesical adverse reactions are rare, clinicians and clinical microbiologists need to be aware of the possibility of disseminated infection by M . bovis BCG in the appropriate setting of clinical history, physical examination, and laboratory investigation. J Clin Microbiol, 1999 Jun, 37(6), 1967 - 70 Comparison of three commercial systems for identification of yeasts commonly isolated in the clinical microbiology laboratory; Wadlin JK et al.; We evaluated three commercial systems (RapID Yeast Plus System; Innovative Diagnostic Systems, Norcross, Ga.; API 20C Aux; bioMerieux-Vitek, Hazelwood, Mo.; and Vitek Yeast Biochemical Card, bioMerieux-Vitek) against an auxinographic and microscopic morphologic reference method for the ability to identify yeasts commonly isolated in our clinical microbiology laboratory . Two-hundred one yeast isolates were compared in the study . The RapID Yeast Plus System was significantly better than either API 20C Aux (193 versus 167 correct identifications; P < 0.0001) or the Vitek Yeast Biochemical Card (193 versus 173 correct identifications; P = 0.003) for obtaining correct identifications to the species level without additional testing . There was no significant difference between results obtained with API 20C Aux and the Vitek Yeast Biochemical Card system (P = 0.39) . The API 20C Aux system did not correctly identify any of the Candida krusei isolates (n = 23) without supplemental testing and accounted for the major differences between the API 20C Aux and RapID Yeast Plus systems . Overall, the RapID Yeast Plus System was easy to use and is a good system for the routine identification of clinically relevant yeasts. J Clin Microbiol, 1999 Jun, 37(6), 1752 - 7 Density and molecular epidemiology of Aspergillus in air and relationship to outbreaks of Aspergillus infection; Leenders AC et al.; After five patients were diagnosed with nosocomial invasive aspergillosis caused by Aspergillus fumigatus and A . flavus, a 14-month surveillance program for pathogenic and nonpathogenic fungal conidia in the air within and outside the University Hospital in Rotterdam (The Netherlands) was begun . A . fumigatus isolates obtained from the Department of Hematology were studied for genetic relatedness by randomly amplified polymorphic DNA (RAPD) analysis . This was repeated with A . fumigatus isolates contaminating culture media in the microbiology laboratory . The density of the conidia of nonpathogenic fungi in the outside air showed a seasonal variation: higher densities were measured during the summer, while lower densities were determined during the fall and winter . Hardly any variation was found in the numbers of Aspergillus conidia . We found decreasing numbers of conidia when comparing air from outside the hospital to that inside the hospital and when comparing open areas within the hospital to the closed department of hematology . The increase in the number of patients with invasive aspergillosis could not be explained by an increase in the number of Aspergillus conidia in the outside air . The short-term presence of A . flavus can only be explained by the presence of a point source, which was probably patient related . Genotyping A . fumigatus isolates from the department of hematology showed that clonally related isolates were persistently present for more than 1 year . Clinical isolates of A . fumigatus obtained during the outbreak period were different from these persistent clones . A . fumigatus isolates contaminating culture media were all genotypically identical, indicating a causative point source . Knowledge of the epidemiology of Aspergillus species is necessary for the development of strategies to prevent invasive aspergillosis . RAPD fingerprinting of Aspergillus isolates can help to determine the cause of an outbreak of invasive aspergillosis. Rev Clin Esp, 1999 Mar, 199(3), 132 - 5 {Primary resistance of Mycobacterium tuberculosis in 10 hospitals of the community of Castille and Leon . Group of Microbiologists from Castille and Leon}; Alberte Castineiras A et al.; OBJECTIVE: To determine the primary drug resistance (PDR) of Mycobacterium tuberculosis in ten hospitals in the Castile-Leon Community for a five-year period (1991-1995), in a sanitary area with almost two millions of inhabitants . MATERIAL AND METHODS: The sensitivity of 825 strains of Mycobacterium tuberculosis to antituberculous drugs was studied using the proportion method; 773 strains were from HIV-negative patients and 52 from HIV-positive patients . RESULTS: Thirty-four out of the 824 strains were resistant to one or more drugs: 31 (4%) from HIV-negative patients and 3 (5.7%) from HIV-positive patients . The resistance to the different drugs for strains from HIV-negative patients was: streptomycin, 2.4%; isoniazid, 1.8%; ethambutol, 0.6%, and rifampin, 0.2% . For HIV-positive patients, resistance to streptomycin was 5.7% and to isoniazid 1.9% . Resistance to a single agent was the resistance mode observed most commonly: 23 (2.9%) in HIV-negative patients and 2 (3.3%) in HIV-positive patients . There was not a single strain resistant to isoniazid and rifampin . CONCLUSIONS: The incidence of PR in the surveyed area was low, including isoniazid . The group of HIV-positive patients did not show a significant increase in resistance (p = 0.4; OR, 1.44; 95% CI, 0.43-4.86) . Regular surveillance of drug resistance is recommended to adjust therapeutic regimes. Trop Gastroenterol, 1998 Oct-Dec, 19(4), 145 - 7 Prevalence of hepatitis B virus infection amongst healthy Nepalese males; Manandhar K et al.; Sera from 478 healthy Nepalese males, inhabiting various districts of five development regions of Nepal were collected from October 1996 to March 1997 and examined for the presence of HBsAg by third generation ELISA and latex agglutination test in the laboratory of Central Department of Microbiology, Tribhuvan University, Kirtipur . The surface antigen was detected by ELISA in 4% (19/478) of subjects studied . However, different results were obtained in Latex agglutination test . The correlation between the results obtained from these two different tests is statistically significant (X2 = 85.11, P < 0.05) . The results obtained from ELISA showed the probability of Nepal to fall in WHO category of intermediate endemicity zone for hepatitis B infection . The percent positivity of HBsAg was found to increase steadily from Eastern (2%) to Far Western (6%) development regions . None of the samples represented from Kathmandu valley were positive for HBsAg . Hepatitis B infection was present in the age groups of 16-20 years and 36-40 years . The levels of ALT were assayed in all samples recorded positive for HBsAg from ELISA . Sixteen percent (3/19) of the infected subjects had raised ALT levels while 84% (16/19) had normal levels. Infect Immun, 1999 May, 67(5), 2266 - 76 Membrane topology and cellular location of the Treponema pallidum glycerophosphodiester phosphodiesterase (GlpQ) ortholog; Shevchenko DV et al.; Recent reports that isolated Treponema pallidum outer membranes contain an ortholog for glycerophosphodiester phosphodiesterase (GlpQ) (D . V . Shevchenko, D . R . Akins, E . J . Robinson, M . Li, O . V . Shevchenko, and J . D . Radolf, Infect . Immun . 65:4179-4189, 1997) and that this protein is a potential opsonic target for T . pallidum (C . E . Stebeck, J . M . Shaffer, T . W . Arroll, S . A . Lukehart, and W . C . Van Voorhis, FEMS Microbiol . Lett . 154:303-310, 1997) prompted a more detailed investigation of its physicochemical properties and cellular location . {14C}palmitate radiolabeling studies of a GlpQ-alkaline phosphatase fusion expressed in Escherichia coli confirmed the prediction from DNA sequencing that the protein is lipid modified . Studies using Triton X-114 phase partitioning revealed that the protein's amphiphilicity is due to lipid modification and that a substantial portion of the polypeptide is associated with the T . pallidum peptidoglycan sacculus . Three different approaches, i.e., (i) proteinase K treatment of intact treponemes, (ii) indirect immunofluorescence analysis of treponemes encapsulated in agarose beads, and (iii) opsonophagocytosis of treponemes incubated with antiserum against recombinant GlpQ by rabbit peritoneal macrophages, confirmed that GlpQ is entirely subsurface in T . pallidum . Moreover, rabbits hyperimmunized with GlpQ were not protected against intradermal challenge with virulent treponemes . Circular dichroism spectroscopy confirmed that the recombinant form of the polypeptide lacked discernible evidence of denaturation . Finally, GlpQ was not radiolabeled when T . pallidum outer membranes were incubated with 3-(trifluoromethyl)-3-(m-{125I}iodophenyl)-diazarene, a photoactivatable, lipophilic probe which promiscuously labels both proteins and lipids within phospholipid bilayers . Taken as a whole, these studies indicate that the T . pallidum GlpQ ortholog is a periplasmic protein associated predominantly with the spirochete's peptidoglycan-cytoplasmic membrane complex. Infect Immun, 1999 May, 67(5), 2233 - 40 Replication of Toxoplasma gondii, but not Trypanosoma cruzi, is regulated in human fibroblasts activated with gamma interferon: requirement of a functional JAK/STAT pathway; Ceravolo IP et al.; To study the role of tryptophan degradation by indoleamine 2, 3-dioxygenase (INDO) in the control of Trypanosoma cruzi or Toxoplasma gondii replication, we used human fibroblasts and a fibrosarcoma cell line (2C4) . The cells were cultured in the presence or absence of recombinant gamma interferon (rIFN-gamma) and/or recombinant tumor necrosis factor alpha (rTNF-alpha) for 24 h and were then infected with either T . cruzi or T . gondii . Intracellular parasite replication was evaluated 24 or 48 h after infection . Treatment with rIFN-gamma and/or rTNF-alpha had no inhibitory effect on T . cruzi replication . In contrast, 54, 73, or 30% inhibition of T . gondii replication was observed in the cells treated with rIFN-gamma alone, rIFN-gamma plus rTNF-alpha, or TNF-alpha alone, respectively . The replication of T . gondii tachyzoites in cytokine-activated cells was restored by the addition of extra tryptophan to the culture medium . Similarly, T . gondii tachyzoites transfected with bacterial tryptophan synthase were not sensitive to the microbiostatic effect of rIFN-gamma . We also investigated the basis of the cytokine effect on parasite replication by using the three mutant cell lines B3, B9, and B10 derived from 2C4 and expressing defective STAT1alpha (signal transducer and activator of transcription), JAK2 (Janus family of cytoplasmic tyrosine kinases), or JAK1, respectively, three important elements of a signaling pathway triggered by rIFN-gamma . We found that rTNF-alpha was able to induce low levels expression of INDO mRNA in the parental cell line, as well as the cell line lacking functional JAK2 . In contrast to the parental cell line (2C4), rIFN-gamma was not able to induce the expression of INDO mRNA or microbiostatic activity in any of the mutant cell lines . These findings indicate the essential requirement of the JAK/STAT pathway for the induction of high levels of INDO mRNA, tryptophan degradation, and the anti-Toxoplasma activity inside human nonprofessional phagocytic cells. Clin Chem, 1999 May, 45(5), 739 - 45 Genetic testing and the clinical laboratory improvement amendments of 1988: present and future; Schwartz MK; CLIA '88 superseded CLIA '67 . CLIA '88 set standards designed to improve quality and expanded federal oversight to virtually all clinical laboratories in the United States . Presumably because genetics testing was then in its infancy, CLIA '88 did not devote a special section to genetics testing . Biochemical and immunochemical tests used to evaluate inborn errors of metabolism and other genetic entities were categorized as analytes in the Clinical Chemistry section, and DNA probes used primarily in infectious disease were included in Microbiology . The legal, social, economic, and ethical implications of genetic testing and the rapid commercialization of these tests led to recommendations that genetic testing be defined as a laboratory specialty with a subsection in CLIA . The advisory committee created under CLIA was assigned to review these recommendations . The committee agreed that genetics testing was sufficiently different from other areas already included in CLIA to warrant a separate section . Two definitions were adopted . The more clear-cut one is for molecular genetic and cytogenic tests . This includes the analysis of human DNA/RNA in evaluating genetic diseases . The second definition is not as clear-cut and is for the analysis of proteins and metabolites used predominantly to detect inborn errors of metabolism . Many of these analytes already are categorized according to their uses for other purposes . The recommendations for genetic testing include detailed and specific proposals concerning personnel, confidentiality and informed consent, quality control, contamination, proficiency testing, validation of tests, special reporting, retention of records, and reuse of tested specimens. Enferm Infecc Microbiol Clin, 1999 Mar, 17(3), 119 - 25 {Evaluation of a simple rapid polymerase chain reaction (PCR) technique for the diagnosis of Helicobacter pylori infection in childhood}; Munoz C et al.; BACKGROUND: The purpose of this study was to evaluate the use of polymerase chain reaction (PCR) applied to the diagnosis Helicobacter pylori infection in the pediatric population, by means of a rapid and simple method of extraction and posterior detection by a colorimetric hybridation of amplified H . pylori DNA . METHODS: Fifty three gastric biopsies, obtained through upper gastrointestinal endoscopy from 53 patients with gastric pathology (45 recidivant abdominal pain and eight hematemesis), were processed from october 1995 to july 1996 . Three non invasive tests were performed: detection of IgG by (Cobas) Core Anti-H . pylori EIA (Roche), breath test with 13C-urea and PCR of dental plaque, as well as four invasive ones: histologic study, culture into selective (Pylori Agar, bioMerieux) and non selective media (Columbia Agar with 5% sheep blood, bioMerieux), test of rapid urease and PCR of gastric biopsies . A 20% solution of Chelex 100 resin (DNA Extraction Reagen, Perkin Elmer) was used for DNA extraction, amplification was performed from gen ureA (Clayton, 1992) and amplified DNA was detected by colorimetric hybridation (PCR ELISA, Boehringer, Mannheim) . Duration of the PCR process was: extraction 25 min, amplification two hours and detection three hours . RESULTS: Results of culture and PCR from gastric biopsies agreed in 84.3% of cases (27 positives, 16 negatives and two not determined) . Two samples were positive by culture and negative by PCR, and were considered as PCR false negatives due to positivity of three or more other tests . Six samples were negative by culture and positive by PCR, being considered as culture false negatives due to positivity of three or more other tests . Sensitivity of PCR and culture was 94.2 and 82.8% . Specificity was 100% for both tests . None of the dental plaque samples was positive . CONCLUSIONS: When invasive techniques are to be done for microbiologic diagnose of H . pylori, PCR increases the confirmation rate of infection; the present procedure enables daily routine work due to its simplicity and its short turnaround time. Enferm Infecc Microbiol Clin, 1999 Mar, 17(3), 113 - 8 {outbreak of invasive pulmonary mycosis in neutropenic hematologic patients in relation to remodelling construction work}; Gaspar C et al.; BACKGROUND: An increase was observed in the number of cases of invasive pulmonary mycosis in neutropenic hematologic patients coinciding with construction work being carried out in the Hospital Clinico San Carlos (Madrid, Spain) . The aim of this study was to confirm the existence of an outbreak, identify the factors related and adopt adequate control measures . METHODS: A descriptive, epidemiologic, environmental microbiologic study was performed . The incidence of cases in the study period and a control period was compared . Air samples were collected in the affected area and other areas of hospitalization not related to the construction work . The ventilation system of the Hemalotogy Isolation Unit (HIU) was also sampled . The control measures undertaken included: hermetic sealing of the construction work adjacent to hematology followed by transfer of the patients to another floor of the hospital . RESULTS: The existence of an outbreak was confirmed (significant increase in the incidence, p = 0.017) . Eleven cases and one death by massive hemoptisis were reported . The mean total fungal count and to Aspergillus spp were 120 and 35 UFC/m3, respectively in the hematologic hospitalization area adjacent to the construction work and 37 and 5 UFC/m3 in other areas (p < 0.001) . Contamination was detected in the ventilation system of the HIU by A . fumigatus (125 UFC/m3 of air from the interior of the conduct) . CONCLUSIONS: An elevated number of fungal spores found in samples from the hematologic hospitalization area as well as no further new cases being reported following the transfer of the patients suggest that the outbreak was related to the adjacent construction work being carried out. Retina, 1999, 19(2), 98 - 102 Microbiologic yields and complication rates of vitreous needle aspiration versus mechanized vitreous biopsy in the Endophthalmitis Vitrectomy Study; Han DP et al.; PURPOSE: To compare the microbiologic yields and complication rates associated with vitreous needle tap and vitreous biopsy in the Endophthalmitis Vitrectomy Study (EVS) . METHODS: Of 420 EVS patients with postoperative endophthalmitis, 201 received immediate vitreous tap or biopsy (without pars plana vitrectomy) by random assignment and 193 completed 9-12 months of follow-up . Vitreous specimens were obtained by biopsy with a 20-gauge vitrectomy cutting instrument or by needle tap with a 22-27-gauge needle . If resistance to aspiration by needle tap was noted, a vitreous biopsy was performed . RESULTS: Of 201 patients undergoing tap or biopsy, 70 (35%) had needle tap, 127 (63%) had mechanized biopsy, and 4 (2%) had initial needle tap that was aborted to mechanized biopsy ("abort" eyes) . Intraoperative hyphema occurred in 2 tap eyes (3%), 3 biopsy eyes (2%), and 0 (0%) abort eyes . Postoperative retinal detachment developed in 8 (11%) tap eyes, 10 (8%) biopsy eyes, and 0 (0%) abort eyes (not significant) . Respective rates of culture and gram stain positivity were 69% and 42% in tap eyes and 66% and 41% in biopsy eyes (not significant) . The rate of severe visual loss (final acuity <5/200) was significantly higher in tap eyes (16 eyes, 24%) compared with biopsy eyes (13 eyes, 11%) and abort eyes (0 eyes, 0%; P = 0.043) . The difference was largely explained by the greater proportion of virulent organisms in the tap eyes compared with biopsy eyes . When visual acuity outcome was defined by other thresholds (20/40 and 20/100), the difference was not significant . CONCLUSIONS: This study showed no significant differences between mechanized vitreous biopsy and needle tap with respect to microbiologic yield, operative complications, short-term (9-12 months) retinal detachment risk, or visual outcome . Choice of vitreous sampling procedure must depend on the clinical judgment of the surgeon. Pathology, 1999 Feb, 31(1), 67 - 9 CSF bacterial antigen detection tests offer no advantage over Gram's stain in the diagnosis of bacterial meningitis; Mein J et al.; While bacterial antigen detection (BAD) tests have been used on cerebrospinal fluid (CSF) with success in the diagnosis of bacterial infection in developing countries, their value in the developed world has been recently questioned . In Darwin, Northern Territory (NT), there are good diagnostic resources but high rates of infectious disease, so it was unclear which findings were applicable to our own population . This study aimed to determine the utility of the BAD tests in detection of bacterial meningitis from CSF in patients studied at Darwin, using a retrospective review of hospital case records and microbiology laboratory reports, over a 19 month period, and utilising a clinical component in the case definition of bacterial meningitis . The sensitivity of the BAD test in the diagnosis of acute bacterial meningitis was 28.6%, with a specificity of 98.7% and a positive predictive value of 85.7% . The cost per positive test was computed at $240 . No cases of bacterial meningitis which were positive on the BAD test were missed on Gram's stain of CSF . We conclude that in our setting BAD tests alone are not sensitive enough to confidently diagnose bacterial meningitis . BAD tests are more costly and offer no advantage in speed of diagnosis or in antibiotic pre-treated patients, compared to routine Gram's stain. Pathology, 1999 Feb, 31(1), 47 - 50 A fluffy white traveller: imported Coccidiodes immitis infection in an Australian tourist; Baird RW et al.; Pulmonary coccidioidomycosis is a rare cause of pulmonary nodules and respiratory infection in travellers to endemic areas . An Australian tourist suffered an acute respiratory illness while on holiday in Mexico . She subsequently developed erythema nodosum and was noted to have a left pulmonary nodule on chest X-ray after return to Australia . The diagnosis of Coccidioides immitis infection was established by histology and culture of the resected lung lesion . The patient made an uneventful recovery and received one month of therapy with ketoconazole . Culture of the fungus took place under controlled Class 3 conditions . An unusual fungal infection in Australia, coccidioidomycosis poses special risks to staff of microbiology laboratories. Am J Vet Res, 1999 Apr, 60(4), 473 - 80 Evaluation of severe disease induced by aerosol inoculation of calves with bovine respiratory syncytial virus; Woolums AR et al.; OBJECTIVE: To develop a model of bovine respiratory syncytial virus (BRSV) infection that induces severe disease similar to that seen in some cattle with naturally acquired BRSV infection . ANIMALS: 25 male Holstein calves, 8 to 16 weeks old . PROCEDURE: 17 calves were given a low-passage field isolate of BRSV by aerosolization; 8 control calves were given supernatant from noninfected cell culture . Disease was characterized by evaluating clinical signs, virus isolation and pulmonary function tests, and results of blood gas analysis, gross and histologic postmortem examination, and microbiologic testing . RESULTS: Cumulative incidence of cough, harsh lung sounds, adventitious sounds, and dyspnea and increases in rectal temperature and respiratory rate were significantly greater in infected calves . Three infected calves developed extreme respiratory distress and were euthanatized 7 days after inoculation . Virus was isolated from nasal swab specimens from all infected calves but not from mock infected calves . On day 7 after inoculation, mean PaO2 and PaCO2 were significantly lower, and pulmonary resistance was significantly higher, in infected calves . During necropsy, infected calves had varying degrees of necrotizing and proliferative bronchiolitis and alveolitis with syncytial formation . The 3 calves euthanatized on day 7 had emphysematous bullae in the caudal lung lobes; 1 had unilateral pneumothorax . CONCLUSION AND CLINICAL RELEVANCE: Severe disease similar to that seen in some cattle with naturally acquired BRSV infection can be induced in calves with a single aerosol exposure of a low-passage clinical isolate of BRSV . Our model will be useful for studying the pathogenesis of BRSV infection and for evaluating vaccines and therapeutics. Infection, 1999 Jan-Feb, 27(1), 63 - 66 Microbiology in intraabdominal infections: what is the message for clinical studies? (Opinion paper); Condon RE; Bacterial flora participating in intraabdominal infections are well known . Data are reviewed here documenting the constant antibiotic susceptibility of this flora despite many years of widespread use of antibiotics . The flaws inherent in the concept of "microbiologic efficacy" in evaluating antibiotic efficacy are examined . Among the possible results of a clinical trial, outcomes not in accord with antibiotic sensitivity data occur regularly . Factors such as host defense responses, source control and inoculum density also influence the outcome in a clinical trial but are not currently being measured . Recommendations for improved conduct of trails are made. J Clin Microbiol, 1999 May, 37(5), 1612 - 6 Pseudoepidemic of Aspergillus niger infections traced to specimen contamination in the microbiology laboratory; Laurel VL et al.; We report a pseudo-outbreak of Aspergillus niger that followed building construction in our clinical microbiology laboratory . Because outbreaks of invasive aspergillosis have been linked to hospital construction, strategies to minimize dust in patient care areas are common practice . We illustrate that the impact of false-positive cultures on patient care should compel laboratories to prevent specimen contamination during construction. Med Mycol, 1999 Feb, 37(1), 11 - 7 Comparison of four commercialized biochemical systems for clinical yeast identification by colour-producing reactions; Paugam A et al.; We compared the ability of four commercially available yeast identification systems for routine laboratory hospital use: Auxacolor (AUX) (Sanofi Diagnostics Pasteur, Marne-la-Coquette), Fungichrom I (FUC) and Fungifast I Twin (FUF) (International Microbio, Toulon), Api Candida (API) (bioMerieux, Lyon) . These systems are based on obtaining a biochemical profile easily defined by colorimetric reactions . We tested 202 yeasts belonging to 19 species which were included or were not included in the manufacturer's data base of the identification systems . Without extra tests, for all the organisms tested, after 24 h of incubation, the percentage of organisms correctly identified was 48% for AUX, 75% for FUC, 77% for FUF and 81% for API . However, if we consider the ratio of the number of correct identifications without extra tests with the number of yeasts included in the manufacturers' data bases (sensitivity) the results increased to 61% for AUX, 81% for FUC, 91% for FUF and 83% for API . These systems are particularly well adapted to medical use, they are simple to set up, interpret, and have very good efficiency for the yeasts most commonly isolated in clinical specimens . The findings reported here indicate that the most favourable results were obtained with FUF and API systems. Vopr Pitan, 1999, 68(1), 46 - 8 {Medical aspect of food starch}; Maksimov VI et al.; In the review are considered aspects of food starch such as its relationship with cancer of the large intestine, its absorption, digestion and metabolism, influence of starch on the composition and biochemical characteristics of microbiocenosis. Hosp Med, 1999 Jan, 60(1), 24 - 8 Gastrointestinal manifestations of HIV infection; Blanshard C; As patients with human immunodeficiency virus (HIV) infection become more immunocompromised, gastrointestinal symptoms become more common . Most symptoms result from opportunistic infections and can be diagnosed and treated by gastroenterologists, although medical microbiology and histopathology input is essential. J CANNT, 1998 Fall, 8(4), 20 - 1 Hepatitis B vaccine as a cause of false positive hepatitis B surface antigen; Olde C et al.; The hemodialysis unit at The Toronto Hospital routinely vaccinates its patients against hepatitis B using Engerix B Hepatitis B Vaccine (Recombinant) . A new positive hepatitis B surface antigen (HBsAg) was flagged by the lab, but upon investigation it was discovered that the patient had received hepatitis vaccine two days earlier . The infectious diseases service was consulted and the case was referred to the microbiologist . Subsequent testing showed a conversion to negative antigen . We decided to test other patients who had just been vaccinated to see if we could replicate our results . Seven other patients were selected, and we found the incidence of false positive HBsAg to be 50% . The duration of the vaccine-induced positive HBsAg lasted no more than two weeks . Our results are compatible with the results found in the current literature. Am J Infect Control, 1999 Apr, 27(2), 158 - 64 Outcomes of a postdischarge surveillance system for surgical site infections at a Midwestern regional referral center hospital; Fields CL; BACKGROUND: Postdischarge surveillance has been reported in the literature as one method for detecting surgical site infections (SSIs) that more traditional methods of surveillance (review of readmission data, monitoring of microbiology, radiology, pharmacy antibiotic usage reports, and medical record review) fail to include . METHODS: This article describes a postdischarge surveillance program that used surgeon questionnaires and was implemented at a 225-bed Midwestern regional referral center hospital . Evaluation of the postdischarge program was accomplished by review of infection control program data for calendar years 1995 through 1997 . RESULTS: Implementation of the postdischarge program resulted in an almost fourfold increase (in both 1995 and 1996) in SSI rates over the reported SSI rates if only traditional surveillance methods had been used . A majority of surgeons (79% in 1995 and 83% in 1996) had individual response rates of 80% or greater . In addition, implementation of the postdischarge program required only 3.5 to 4 additional hours per month . CONCLUSIONS: Results suggest that the postdischarge surveillance program identified SSIs missed by traditional surveillance methods, resulted in higher reported rates of SSI, was moderately to well accepted by surgeons, and was implemented with a minimum of organizational resources. Am J Infect Control, 1999 Apr, 27(2), 97 - 132; quiz 133-4; discussion 96 Guideline for Prevention of Surgical Site Infection, 1999 . Centers for Disease Control and Prevention (CDC) Hospital Infection Control Practices Advisory Committee; Mangram AJ et al.; EXECUTIVE SUMMARY The "Guideline for Prevention of Surgical Site Infection, 1999" presents the Centers for Disease Control and Prevention (CDC)'s recommendations for the prevention of surgical site infections (SSIs), formerly called surgical wound infections . This two-part guideline updates and replaces previous guidelines.1,2 Part I, "Surgical Site Infection: An Overview," describes the epidemiology, definitions, microbiology, pathogenesis, and surveillance of SSIs . Included is a detailed discussion of the pre-, intra-, and postoperative issues relevant to SSI genesis.Part II, "Recommendations for Prevention of Surgical Site Infection," represents the consensus of the Hospital Infection Control Practices Advisory Committee (HICPAC) regarding strategies for the prevention of SSIs.3 Whenever possible, the recommendations in Part II are based on data from well-designed scientific studies . However, there are a limited number of studies that clearly validate risk factors and prevention measures for SSI . By necessity, available studies have often been conducted in narrowly defined patient populations or for specific kinds of operations, making generalization of their findings to all specialties and types of operations potentially problematic . This is especially true regarding the implementation of SSI prevention measures . Finally, some of the infection control practices routinely used by surgical teams cannot be rigorously studied for ethical or logistical reasons (e.g., wearing vs not wearing gloves) . Thus, some of the recommendations in Part II are based on a strong theoretical rationale and suggestive evidence in the absence of confirmatory scientific knowledge.It has been estimated that approximately 75% of all operations in the United States will be performed in "ambulatory," "same-day," or "outpatient" operating rooms by the turn of the century.4 In recommending various SSI prevention methods, this document makes no distinction between surgical care delivered in such settings and that provided in conventional inpatient operating rooms . This document is primarily intended for use by surgeons, operating room nurses, postoperative inpatient and clinic nurses, infection control professionals, anesthesiologists, healthcare epidemiologists, and other personnel directly responsible for the prevention of nosocomial infections.This document does not: Specifically address issues unique to burns, trauma, transplant procedures, or transmission of bloodborne pathogens from healthcare worker to patient, nor does it specifically address details of SSI prevention in pediatric surgical practice . It has been recently shown in a multicenter study of pediatric surgical patients that characteristics related to the operations are more important than those related to the physiologic status of the patients.5 In general, all SSI prevention measures effective in adult surgical care are indicated in pediatric surgical care . Specifically address procedures performed outside of the operating room (e.g., endoscopic procedures), nor does it provide guidance for infection prevention for invasive procedures such as cardiac catheterization or interventional radiology . Nonetheless, it is likely that many SSI prevention strategies also could be applied or adapted to reduce infectious complications associated with these procedures . Specifically recommend SSI prevention methods unique to minimally invasive operations (i.e., laparoscopic surgery) . Available SSI surveillance data indicate that laparoscopic operations generally have a lower or comparable SSI risk when contrasted to open operations.6-11 SSI prevention measures applicable in open operations (e.g., open cholecystectomy) are indicated for their laparoscopic counterparts (e.g., laparoscopic cholecystectomy) . Recommend specific antiseptic agents for patient preoperative skin preparations or for healthcare worker hand/forearm antisepsis . Hospitals should choose from products recommended for these activitie Clin Microbiol Rev, 1999 Apr, 12(2), 183 - 6 Clinical Microbiology Reviews: genesis of a journal; Morello JA; In 1986 planning for a new ASM review journal, Clinical Microbiology Reviews (CMR), began . CMR would publish articles primarily of interest to persons concerned with pathogenesis, laboratory diagnosis, epidemiology, and control of human and veterinary pathogens . The first issue was published in January 1988, with quarterly publication since then . The journal quickly became successful in terms of subscribers and impact on the field, earning a strong national and international reputation . The achievements of CMR are owed to many persons, including the editorial board, the production team, and especially the contributing authors. Am J Cardiol, 1999 Apr 1, 83(7), 1075 - 9 Clinical course, microbiologic profile, and diagnosis of periannular complications in prosthetic valve endocarditis; San Roman JA et al.; Whether periannular extension of prosthetic valve endocarditis (abscesses, pseudoaneurysms, fistulas) is related to the etiologic agent, the clinical course and the prognosis is still unknown . Likewise, transesophageal echocardiographic accuracy in detecting periannular complications in prosthetic recipients remains unsettled . We retrospectively analyzed data from 87 patients with anatomically proven prosthetic valve endocarditis who underwent a transesophageal echocardiographic examination . Periannular complications (30 abscesses, 18 pseudoaneurysms, 8 fistulas; 8 with >1) were found in 46 patients; results were compared with the remaining 41 without complications . Transesophageal echocardiography correctly identified 27 abscesses (90%) and all pseudoaneurysms and fistulas . One diagnosis of pseudoaneurysm by echocardiography was not found at surgery . No statistical differences were found regarding age, sex, type of prosthesis (mechanical vs biologic), and etiologic agent . Periannular complications were more frequent in aortic location (70% vs 20% in mitral position; p <0.001) and in early (within 6 months after surgery) endocarditis (63% vs 38% in late endocarditis; p = 0.04) . The same percentage of patients from both groups underwent surgery (98% with and 90% without complications) . At discharge, 62% and 67% of patients were alive, respectively . Thus, periannular complications in prosthetic valve endocarditis are more frequent in aortic location and within 6 months after surgery . Neither the type of prosthesis nor the etiologic agent are related to the presence of periannular complications . Short-term prognosis in patients who underwent surgery is not affected by the presence of periannular complications. Can J Hosp Pharm, 1994 Jun, 47(3), 111 - 6 The impact of formulary reservations on drug utilization: a controlled trial; Mather JL et al.; A controlled trial was conducted in two teaching hospitals (A and B), with similar case mixes to determine the impact of reservations, which were educational in nature, on the utilization of oral ciprofloxacin . Over a two-month period the health records of all the patients who received the drug were reviewed, and information on utilization and demographics of patients receiving the drug was recorded . As well, the number of admissions to the two hospitals over this period were compared . If culture and sensitivity (C & S) results were available, appropriateness was assessed in accordance with criteria for use established at site A; in the absence of C & S information, consensus by two microbiologists was used . Over the two-month period a total of 136 patients received ciprofloxacin at the two institutions . At site A, which had reservations, the number of patients who continued to receive ciprofloxacin upon admission was significantly decreased relative to site B, which did not have reservations (14% vs . 36% respectively, p = .029) . As well, when assessed by total number of admissions to the institutions, the number of patients receiving ciprofloxacin at site A was less than site B (1.5% vs . 2.6% respectively, p = .003)) . While the utilization was decreased at site A vs . site B, the proportion of patients with therapy deemed to be appropriate was not different between the two sites . Educationally based reservations are an effective formulary tool for optimizing drug utilization. Hosp Formul, 1993 Jan, 28 Suppl 1, 55 - 8 The true cost of monitoring antibiotic levels; Gruneberg RN; Antibiotic assays are most often ordered for the purpose of toxicity monitoring, which usually involves determination of peak and trough antibiotic concentrations in the blood . The cost of monitoring antibiotic levels is probably higher than is commonly appreciated . Factors that contribute to the cost of this service include staffing the microbiology laboratory with appropriate personnel, who are responsible for determining the adequacy of sample collection and related patient information; analyzing the specimens in a timely manner; and taking action to modify drug dosage and dosage intervals in light of the test results . There are also costs related to the reagents, consumables, and equipment used in the assay, as well as to revenue and capital overheads . Additional clinical and laboratory costs can be incurred in the event of litigation pursuant to antibiotic-induced toxicity . With hospital and government policymakers devoting increasing attention to the escalating costs of health care, pressure to move away from the routine use of drugs having dose-related toxicity may increase, the objective being to save on assay costs. Hosp Pharm, 1992 Jul, 27(7), 596 - 603, 614 Department of pharmacy-initiated program for streamlining empirical antibiotic therapy; Pastel DA et al.; The outcome of a department of pharmacy-initiated "streamlining" study designed to promote cost-conscious modifications of empirically selected antibiotic therapy is described . Two hundred forty-one evaluable adult patients started on restricted-use antibiotics at this university-affiliated community private teaching hospital were enrolled in a 9-week prospective streamlining study . Patients were alternately assigned to a Control (i.e., no pharmacist-initiated streamlining recommendations offered based on culture and susceptibility reports) or a Pharmacist Intervention group (i.e., pharmacist offers recommendations to streamline therapy) . A statistically significant greater number of patients had their empiric antibiotic treatment courses modified to more appropriate antibiotic choices after receipt of culture and susceptibility reports among private prescribers in the Pharmacist Intervention group (83%) than in the Control group (38%) (p = .006) . Additionally, pharmacists were overall successful in gaining prescriber acceptance for 64% of recommended changes of empiric antibiotic treatment courses before the receipt of culture and susceptibility reports (e.g., dose and/or frequency changes) . There was no program effect observed with respect to improved physician response to microbiologic data that would allow streamlining empirical antibiotic choices in the Housestaff (i.e., medical or surgical residents), or infectious disease consultant prescriber groups . Projected overall annual cost savings that would be achieved as a result of continued efforts by pharmacists directed at streamlining empirical "restricted" antibiotic regimens is approximately +40,000. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1999 Mar, 87(3), 357 - 61 Massive osteolysis of the mandible: report of a case with multifocal bone loss; Bouloux GF et al.; Osteolysis of the jaws has been reported in association with infection, cysts, neoplasia, and metabolic, endocrine, or hematologic abnormalities . Rare cases of idiopathic osteolysis have also been recorded . We report the case of a 10-year-old girl with mandibular basal and alveolar bone resorption that has continued over a period of 9 years . The patient has subsequently developed bilateral resorption of the ascending rami and condyles . The maxilla is uninvolved . Investigations included radiology, computerized tomography, scintigraphy, hematology, serum chemistry, endocrinology, histopathology, microbiology, and immunology . Neutrophil chemotaxis, chemiluminescence, and random migration values were low but within the normal range . These findings are interpreted as indicating an unusual variant of massive osteolysis. Ir J Med Sci, 1999 Jan-Mar, 168(1), 21 - 4 Farmer's lung in Ireland (1983-1996) remains at a constant level; McGrath DS et al.; A prospective study was undertaken by the Departments of Respiratory Medicine and Medical Microbiology at the Cork University Hospital, a . to investigate the epidemiology of Farmer's Lung (F.L.) in the Republic of Ireland (pop . 3.5 million), with special reference to the South Western Region of this country (pop . 536,000) and b . to assess any relationship between the prevalence/incidence of F.L . with climatic factors in South West Ireland, between 1983 and 1996 . F.L . incidence remained constant throughout the 13 yrs studied both on a national and a regional basis . A significant relationship was also found between total rainfall each summer and F.L . incidence and prevalence over the following yr (p < 0.005) in South-West Ireland . The persistence of F.L . in Ireland at a constant level suggests that farmers' working environment and farm practices need to be improved. Yonsei Med J, 1998 Dec, 39(6), 485 - 7 Surveillance of resistant pathogens and rational use of antibiotics: general remarks; Acar JF; Surveillance of resistant pathogens should lead to improved treatment of patients and to a rational use of antibiotics . The process for decision making between microbiology, general practice and health policy is still to be documented with careful studies. Int J Tuberc Lung Dis, 1999 Feb, 3(2), 149 - 52 Drug-resistant strains of Mycobacterium tuberculosis isolated in Russia; Stepanshina VN et al.; SETTING: State Research Center for Applied Microbiology, Russian Research Institute of Phthisiopulmonology (Ministry of Health, Moscow) . OBJECTIVE: To analyze drug-resistant clinical isolates of Mycobacterium tuberculosis obtained from patients referred to the institute from different parts of Russia, and to study the mechanisms of their rifampicin resistance . DESIGN: Fifty clinical isolates of M . tuberculosis were analysed . Polymerase chain reaction (PCR) and sequencing were used to study the mechanisms of rifampicin resistance in 25 isolates . RESULTS: Among cultures isolated from 50 patients, drug resistance was detected in 33 . Most of the isolates were resistant to rifampicin (25 isolates), isoniazid (14 isolates), and streptomycin (seven isolates) . Only 6% of the isolates were resistant to one drug, while 14% were resistant to two, 32% to three, 40% to four, and 8% to five drugs . Susceptible isolates were derived from 17 patients . The following point mutations and deletions in the rpoB locus, responsible for high level rifampicin resistance (more than 50 microg/ml in egg-based medium), were detected: G-->A/395 (Arg-->Gln), C-->T/232 (His-->Tyr), C-->T/221 (Ser-->Leu), G-->T/202 (Asp-->Tyr), GA-->TT/202-203 (Asp-->Phe), deltaATGGACCAG/199-207 (Met, Asp, Gin), A-->T/91 (Met-->Leu), TG-->CC/227-228 (Leu-->Ser), GAG-->AGT/349-350-351 (Gln-->Ser), deltaGGG/354(Gly) . CONCLUSION: A number of previously unrecognised genetic modifications in the rpoB region were found in rifampicin-resistant strains isolated from patients from different parts of Russia. Microb Pathog, 1999 Apr, 26(4), 207 - 19 Cloning, expression and significance of MPT53 for identification of secreted proteins of Mycobacterium tuberculosis; Wiker HG et al.; Based on our N -terminal amino acid sequence of MPT53 and a deduced DNA sequence, we searched for the corresponding gene in the Mycobacterium tuberculosis genomic sequence at the Sanger centre, localizing mpt53 close to mpt70 and mpt83 . The gene was cloned and expressed, followed by purification of MPT53 to homogeneity from recombinant M . smegmatis culture fluid . In MPT53 there is 60 % identity with the active site of thioredoxin of M . tuberculosis (MPT46) with two cysteins in a CXXC motif, but MPT53 could not serve as an alternative substrate for thioredoxin reductase . Testing for IgM and IgG1 anti-MPT53 in cattle sera showed that MPT53 is immunogenic following natural and experimental infection with M . bovis . Cloning of mpt53 represents cloning of the last of the 10 proteins originally defined as "secreted proteins" of M . tuberculosis and M . bovis based on determination of their "Localization index" (LI) (J Gen Microbiol 1991;137 : 875-84) . T