Patient-reported complications associated with Campylobacter jejuni infection.

This study aimed to investigate the occurrence of complications, especially musculoskeletal symptoms, after sporadic Campylobacter jejuni enteritis of domestic origin in Finland. This multi-centre cross-sectional study was conducted during a seasonal peak in 2002. Questionnaires were sent to Campylobacter-positive patients, representing different geographical areas, 2 months after collection of positive stool samples. Medical records were viewed in several cases. Besides antimicrobial susceptibility testing C. jejuni isolates were serotyped. A total of 235 patients (58%) returned the questionnaire and 201 C. jejuni-positive patients were finally included in the study. Musculoskeletal symptoms associated with C. jejuni enteritis were frequent (39%); joint pain was most commonly reported (81%). The incidence of reactive arthritis was 4% and that of Achilles enthesopathy and/or heel pain was 9%. Stomach ache during enteritis was associated with the later development of joint pain. Antimicrobial treatment was common but did not prevent complications.

Strain and host characteristics of Campylobacter jejuni infections in Finland.

The relative importance of different risk-factors for Campylobacter infections and the role of bacterial strain and host characteristics are uncertain. Swimming in natural sources of water was recently described as a novel independent risk-factor for domestically-acquired Campylobacter infections. The present study investigated exposure factors and demographical characteristics (collected in a questionnaire), and determined whether Campylobacter jejuni serotypes could be linked to each other or to the severity of the disease in domestically-acquired sporadic C. jejuni infections during a seasonal peak in Finland. Swimming was associated positively with an age of

Non-tuberculous mycobacteria: patterns of isolation. A multi-country retrospective survey.

OBJECTIVE: To collect data on non-tuberculous mycobacteria (NTM) isolated from clinical laboratories in different countries to establish: 1) whether the isolation of NTM was increasing, 2) which species were increasing, and 3) whether there was any pattern of geographical distribution. DESIGN: In 1996, the Working Group of the Bacteriology and Immunology Section of the International Union Against Tuberculosis and Lung Disease contacted 50 laboratories in different countries for the necessary information. RESULTS: The number of patients reported with NTM was 36099 from 14 countries. Mycobacterium avium complex, M. gordonae, M. xenopi, M. kansasii and M. fortuitum were the five species most frequently isolated. There was a significant upward trend for M. avium complex and M. xenopi. Pigmented mycobacteria predominated in Belgium, the Czech Republic and the Mediterranean coast of Spain. Non-chromogenic mycobacteria were found to be predominant in the area of the Atlantic coast of Brazil and in Turkey, the United Kingdom, Finland and Denmark. CONCLUSIONS: There was an increase in the number of NTM isolated from clinical samples of patients. Isolation of the most frequent species is constantly changing in most of the geographical areas, and newer species are emerging due to better diagnostic techniques to detect and identify NTM.

Detection and typing of Campylobacter jejuni and Campylobacter coli and analysis of indicator organisms in three waterborne outbreaks in Finland.

Waterborne outbreaks associated with contamination of drinking water by Campylobacter jejuni are rather common in the Nordic countries Sweden, Norway, and Finland, where in sparsely populated districts groundwater is commonly used without disinfection. Campylobacters, Escherichia coli, or other coliforms have rarely been detected in potential sources. We studied three waterborne outbreaks in Finland caused by C. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacters and sample volumes of 1 to 5,000 ml for analysis of coliforms and E. coli, depending on the sampling site. Multiple samples obtained from possible sources (water distribution systems and environmental water sources) and the use of large sample volumes (several liters) increased the chance of detecting the pathogen C. jejuni in water. Filtration of a large volume (1,000 to 2,000 ml) also increased the rate of detection of coliforms and E. coli. To confirm the association between drinking water contamination and illness, a combination of Penner serotyping and pulsed-field gel electrophoresis (digestion with SmaI and KpnI) was found to be useful. This combination reliably verified similarity or dissimilarity of C. jejuni isolates from patient samples, from drinking water, and from other environmental sources, thus confirming the likely reservoir of an outbreak.

Recurrent Sphingomonas paucimobilis -bacteraemia associated with a multi-bacterial water-borne epidemic among neutropenic patients.

A cluster of septicaemias due to several water-related species occurred in a haematological unit of a university hospital. In recurrent septicaemias of a leukaemic patient caused by Sphingomonas paucimobilis, genotyping of the blood isolates by use of random amplified polymorphic DNA-analysis verified the presence of two distinct S. paucimobilis strains during two of the separate episodes. A strain of S. paucimobilis identical to one of the patient's was isolated from tap water collected in the haematological unit. Thus S. paucimobilis present in blood cultures was directly linked to bacterial colonization of the hospital water system. Heterogeneous finger-printing patterns among the clinical and environmental isolates indicated the distribution of a variety of S. paucimobilis clones in the hospital environment. This link also explained the multi-microbial nature of the outbreak.

Nosocomial Legionella pneumophila serogroup 5 outbreak associated with persistent colonization of a hospital water system.

An outbreak of infections caused by Legionella pneumophila serogroup 5 was detected in a university hospital, and nosocomial reservoirs of the legionella epidemic were examined. Clinical isolates from two patients who had been affected by the L. pneumophila serogroup 5 outbreak, and from another patient with a legionella infection caused by the same serogroup 3 years later, were compared to L. pneumophila serogroup 5 isolates from the hospital water supply by two molecular methods, amplified fragment length polymorphism (AFLP) analysis and random amplified polymorphic DNA analysis (RAPD). Genotyping confirmed the epidemiological linkage of the first two patients, and linked their infections with the hospital water supply. The third clinical strain, which was also linked to the hospital water, was very similar to the epidemic strain. Even though the water distribution system was sanitized (superheat and flush sanitation), the epidemic strain was shown to be persisting in the hospital water outlets several years after its initial discovery.

Comparison of mycobacteria-induced cytotoxicity and inflammatory responses in human and mouse cell lines.

Environmental mycobacteria, which are ubiquitous in nature, are also detected in moisture-damaged buildings. Their potential role inducing the adverse health effects associated with living in moisture damaged buildings requires clarification. To establish a model for these studies, we evaluated inflammatory responsiveness in different cell lines exposed to environmental mycobacterial species. Four mycobacterial isolates belonging to Mycobacterium avium complex and Mycobacterium terrae, recovered from the indoor air sampled when a moldy building was being demolished, were studied for their cytotoxicity and ability to stimulate the production of inflammatory mediators in mouse RAW264.7 and human 28SC macrophage cell lines, and human A549 lung epithelial cell line. Lipopolysaccharide (LPS) was used as a positive control. Production of cytokines (tumor necrosis factor alpha, TNF-alpha; interleukin 6, IL-6; and interleukin beta, IL-1beta) was analyzed immunochemically, nitric oxide (NO) by the Griess method, expression of inducible NO synthase with Western blot analysis, and cytotoxicity with the MTT test. Both human and mouse cells produced NO and IL-6 after mycobacterial exposure. Mouse macrophages also showed production of TNF-alpha induced by both mycobacteria and LPS, whereas the human cell lines failed to produce TNF-alpha after mycobacterial exposure and the human epithelial cell line also failed to respond to LPS. Similarly, only mouse macrophages produced IL-1beta. Mycobacterial exposure was not cytotoxic to human cells and was only slightly cytotoxic to mouse macrophages. The results indicate that environmental mycobacterial isolates from moldy buildings are capable of activating inflammatory mechanisms in both human and murine cells. The human and mouse cell lines, however, differ significantly in the grade and type of the responses.

Pulmonary infection caused by an unusual, slowly growing nontuberculous Mycobacterium.

Mycobacterium triplex, a recently described slowly growing nontuberculous mycobacterium, was isolated from a Finnish patient with pulmonary mycobacteriosis. The disease was successfully treated with antimycobacterial drugs. The strain isolated, which was similar to the type strain but differed slightly from the species description, was regarded as a variant of M. triplex sensu stricto. According to present knowledge this variant of the species has never been isolated before.

Tuberculosis as an occupational hazard for health care workers in Estonia.

SETTING: Tuberculosis incidence has been increasing in the Baltic states since the 1990s, accompanied by the emergence of drug resistance, including multidrug resistance (MDR). In this changing situation, the potential threat of nosocomial spread of tuberculosis to other patients and health care workers (HCW) has remained unrecognised. OBJECTIVE: To investigate the risk of tuberculosis in health care workers in Estonia. DESIGN: Cases of tuberculosis registered among HCWs from 1994 to 1998 were evaluated. The case records were analysed retrospectively and combined with bacteriological data including data on drug resistance. RESULTS: Sixty-seven HCWs (23 physicians, 23 nurses and seven laboratory technicians, 12 assistant nurses and two cleaners), all of whom tested negative for human immunodeficiency virus, were diagnosed as having active tuberculosis. The incidence of tuberculosis among HCWs (mean 91/100,000/year) was 1.5 to three times higher than in the general population. In a chest hospital in charge of regional tuberculosis care, the incidence was 30 to 90 times higher, and was highest among physicians. In 49 HCWs tuberculosis was confirmed by culture. Among these, drug resistance was detected in 23 (49%), 18 (38%) of whom had MDR tuberculosis. CONCLUSIONS: Health care workers, especially those working in a chest hospital where tuberculosis patients were treated, were found to be at an elevated risk of tuberculosis. MDR tuberculosis poses a particular threat which is difficult to combat.

Characterization of Mycobacterium bohemicum isolated from human, veterinary, and environmental sources.

Chemotaxonomic and genetic properties were determined for 14 mycobacterial isolates identified as members of a newly described species Mycobacterium bohemicum. The isolates recovered from clinical, veterinary, and environmental sources were compared for lipid composition, biochemical test results, and sequencing of the 16S ribosomal DNA (rDNA) and the 16S-23S rDNA internal transcribed spacer (ITS) regions. The isolates had a lipid composition that was different from those of other known species. Though the isolates formed a distinct entity, some variations were detected in the features analyzed. Combined results of the phenotypic and genotypic analyses were used to group the isolates into three clusters. The major cluster (cluster A), very homogenous in all respects, comprised the M. bohemicum type strain, nine clinical and veterinary isolates, and two of the five environmental isolates. Three other environmental isolates displayed an insertion of 14 nucleotides in the ITS region; they also differed from cluster A in fatty alcohol composition and produced a positive result in the Tween 80 hydrolysis test. Among these three, two isolates were identical (cluster B), but one isolate (cluster C) had a unique high-performance liquid chromatography profile, and its gas liquid chromatography profile lacked 2-octadecanol, which was present in all other isolates analyzed. Thus, sequence variation in the 16S-23S ITS region was associated with interesting variations in lipid composition. Two of the isolates analyzed were regarded as potential inducers of human or veterinary infections. Each of the environmental isolates, all of which were unrelated to the cases presented, was cultured from the water of a different stream. Hence, natural waters are potential reservoirs of M. bohemicum.

Recurrence of urinary tract infections in adult patients with community-acquired pyelonephritis caused by E. coli: a 1-year follow-up.

In a prospective study, 42 women were followed for recurrence of urinary tract infections (UTIs) for 1 y after an index episode of community-acquired pyelonephritis caused by Escherichia coli. Altogether, 26 repeat episodes were detected. Of these, 20 occurred at least 1 month after the index episode and were regarded as recurrences. In all, 40%, (17 of 42) of the women had recurrences. An earlier history of UTI increased the risk of recurrence: 52%, of the 29 women with previous UTI had at least 1 recurrence, compared with 15%, of the 13 patients without previous UTI. E. coli caused the majority (73%) of the recurrences. Genotype comparisons by RAPD-PCR analysis between E. coli isolates from a patient showed that 75%. of the original and recurrent strains were genetically non-identical. Of the 54 E. coli strains, 42 were carrying genes coding for G adhesins of P fimbriae: 40 isolates carried class II, I class III and 1 carried both class II and III G adhesin genes. Each of the virulence-associated factors (genes for G adhesins, MRHA, haemolysin, type 1C fimbriae, and O and K antigens) was evenly distributed among E. coli isolates of index episodes, independent of the recurrences. The index isolates, however, had more virulence-associated factors than did the isolates from the recurrences which were mainly due to lower UTIs.

Intrauterine inflammation at term: a histopathologic study.

This study was undertaken to examine the presence of inflammation of the uterine and gestational tissues as defined by histopathology in clinically noninfected women with term gestation and intact fetal membranes and to evaluate its correlation with measured clinical variables and neonatal and maternal clinical outcome. Two hundred sixteen clinically noninfected term parturients who underwent cesarean section with intact membranes were analyzed for the presence of inflammatory lesions of the gestational tissues and uterus. Nine hundred eighty-one histologic samples were studied, including 212 samples from both chorion membranes and umbilical cords, and 209 placental, 192 myometrial, and 156 decidual samples. In 208 (96%) cases, either amniotic fluid (AF) or endometrial swab samples were cultivated for bacteria. In 148 (69%) cases, the AF leukocyte count was analyzed by the Gram stain method, and in 77 (36%), AF leukocyte esterase activity (LEA) was evaluated. Leukocytic infiltrations were present in samples from 41 (19%) women, varying from 2% to 10% in the different anatomic sites examined. However, after onset of labor, low-grade decidual inflammation was observed in 29% of cases. Cervical dilation (odds ratio, 4.7; 95% confidence interval, 2.4 to 9.3; P < .00003) and meconium-stained AF at the operation (odds ratio, 5.3; 95% confidence interval, 2.2 to 12.5; P < .00015) were associated with the histologic inflammatory lesions observed in decidual samples, independently of AF or endometrial microbial detection, AF leukocytes, or LEA.

Low virulence of Escherichia coli strains causing urinary tract infection in renal disease patients.

The distribution of urinary bacterial species was determined and the virulence factors of Escherichia coli urinary strains analysed by molecular and phenotyping methods in episodes of urinary tract infection in renal disease patients (n =68) in comparison with other immunocompromised patients (n =59) and non-immunocompromised patients (n =21). Escherichia coli was isolated in 116 (78%) of the 148 patients, being the species most frequently isolated in all groups (75% of renal disease patients, 76% of other immunocompromised patients, 95% of non-immunocompromised patients). All other pathogens showed a similar distribution in the renal disease and other immunocompromised patient groups. All virulence factors of Escherichia coli tested for (genes for G adhesins, expression of MR adhesins, production of haemolysin, presence of certain O and K antigens) were found more often in non-immunocompromised than in immunocompromised patients. The factors allowing the highest degree of discrimination between immunocompromised and non-immunocompromised patients were the prevalence of genes for G adhesins (35% vs. 65%) and expression of MR adhesins (32% vs. 55%). It is concluded that there is a lower prevalence of G adhesins and MR adhesins in Escherichia coli strains from immunocompromised patients than non-immunocompromised patients, suggesting that less virulent Escherichia coli strains may cause urinary tract infections more frequently in renal disease patients and other immunocompromised patients. Moreover, the spectrum of urinary pathogens other than Escherichia coli is similar in both immunocompromised patient groups investigated.

Mycobacterium xenopi and related organisms isolated from stream waters in Finland and description of Mycobacterium botniense sp. nov.

Three scotochromogenic Mycobacterium xenopi-like organisms were isolated from stream waters in Finland. These strains grew at 36-50 degrees C but not at 30 degrees C. One of the three strains was fully compatible with the M. xenopi type strain according to GLC-MS, biochemical tests, and 16S rDNA and 16S-23S rDNA internal transcribed spacer (ITS) sequencing. Two of the strains closely resembled M. xenopi in lipid analyses and biochemical tests, but analysis by GLC-MS verified the presence of two new marker fatty acids (2,4,6,x-tetramethyl-eicosanoic acid and 2,4,6,x,x-pentamethyl-docosanoic acid). The 16S rDNA and ITS region sequences of these two strains differed from those of M. xenopi and other previously described mycobacterial sequences. Therefore, the strains are regarded as new species of slow-growing mycobacteria, for which the name Mycobacterium botniense sp. nov. is proposed. The chemical, physical and microbiological quality of the water reservoirs of M. xenopi and M. botniense are described. As far as is known, this is the first time that M. xenopi has been isolated from natural waters. The strains of M. botniense sp. nov. (E347T and E43) have been deposited in the ATCC as strains 700701T and 700702, respectively.

Accelerated detection and identification of mycobacteria with MGIT 960 and COBAS AMPLICOR systems.

An automated cultivation system for mycobacteria, the MGIT 960 system (MGIT system), was compared in the clinical routine with two variants of Löwenstein-Jensen (L-J) medium. A total of 152 isolates were recovered from 2,015 specimens: 139 (91%) with the MGIT system and 127 (84%) with L-J media (P = 0.05). These included 68 isolates of Mycobacterium tuberculosis, of which 88% grew in the MGIT system and 93% grew in L-J media (P = 0.389), and 84 isolates of mycobacteria other than M. tuberculosis (MOTT), of which 94% grew in the MGIT system and 76% grew in L-J media (P = 0.003). More M. avium complex isolates were detected in the MGIT system (n = 65) than in L-J media (n = 50) (P = 0.001). Growth in the MGIT system was detected in 2 weeks for 78% of the isolates, whereas growth was detected in the two L-J media for 17 and 25% of the isolates, respectively. The mean times to detection of M. tuberculosis were 12 days in the MGIT system and 20 days in L-J media, and for M. avium complex the mean times to detection were 8 and 22 to 25 days, respectively. The contamination rates were similar (8.7 to 8.9%) in all media. A commercial amplification system (COBAS AMPLICOR) was evaluated for its ability to rapidly identify M. tuberculosis, M. avium, and M. intracellulare directly from 393 samples in MGIT system broth. A correct PCR result, as evaluated by culture or clinical data, was obtained for 96% of the samples, with inhibition being detected for 2% of the samples. Of the 89 results positive for M. tuberculosis, 91% were regarded as true positive, 8% were regarded as inconclusive, and 2% were considered false positive. For results positive for M. avium and M. intracellulare, 97 and 79%, respectively, were regarded as true positive. Increased rapidity and enhanced isolation of MOTT were obtained with the MGIT system. COBAS AMPLICOR was suitable for rapid identification of these three common pathogens from MGIT system broth.

Development of tuberculin reactivity and sensitization to M. scrofulaceum and M. fortuitum in children BCG-vaccinated at birth.

Since the incidence of tuberculosis is steadily declining in Finland and infections by environmental mycobacteria may be increasing, the aim of the present study was to evaluate the development of tuberculin reactivity and sensitization to environmental mycobacteria. Healthy Finnish schoolchildren aged 10.4-12.4 yrs (n=201) were tested with tuberculin purified protein derivative RT23, Mycobacterium scrofulaceum RS95 and M. fortuitum RS20 sensitins. The same children had been previously tested with the same antigens and methods at the age of 4-6 yrs in 1989. Rapid waning of tuberculin reactivity and decrease in sensitization to environmental mycobacteria were observed between 4-6 yrs. Both tuberculin and sensitin skin reaction sizes decreased significantly over the 6-yrs period. The mean tuberculin skin reaction size was 3.2 mm in diameter, which was significantly (p<0.001) smaller than the mean induration size (4.8 mm) at the age of 4-6 yrs. Similarly, the mean skin reaction sizes to M. scrofulaceum and M. fortuitum sensitins were 3.4 and 1.7 mm, respectively, which were significantly (p<0.001) smaller than 6 yrs earlier (mean 4.5 and 3.1 mm). The number of zero reactions to all antigens increased significantly during the follow-up period. Contacts with pets or farm animals were associated with larger reactions. In contrast, children suffering from allergic symptoms had smaller reactions. Contacts with mycobacteria, either with Mycobacterium tuberculosis or environmental mycobacteria, seem to be too rare to maintain tuberculin responsiveness and a high sensitivity to other mycobacteria. Different bacille Calmette-Guérin vaccine products and dosages used, the declining incidence of tuberculosis and geographical factors, which can influence environmental mycobacterial exposure, may explain the disparity between the present and previous Finnish studies.

Hospital water supply as a source of disseminated Mycobacterium fortuitum infection in a leukemia patient.

Nosocomial acquisition of Mycobacterium fortuitum led to a disseminated infection in a leukemia patient. A linkage to showerhead water was supported by molecular typing of clinical and environmental isolates. Contamination of the hospital water system with microbes that are relatively resistant to common sanitation processes poses an increased risk of infection to neutropenic patients.

Isolation of mycobacteria from indoor swimming pools in Finland.

The presence of mycobacteria in seven indoor pools in Finland was evaluated by multiple culture methods. Replicate samples, with and without inactivation of chlorine by sodium thiosulfate, were cultured in two laboratories. Laboratory I used two methods: (A) no decontamination and (B) cetylpyridinium chloride (0.005%, 20 min); and Laboratory II two methods: (C) cetylpyridinium chloride (0.005%, 18 h) and (D) oxalic acid (5%, 15 min). Samples processed by methods (A) and (B) were cultured on different egg media of pH 6.3 or 5.8; by method (C) on Middlebrook and Cohn 7H10 (+OADC) agar of pH 5.5; and by method (D) on Middlebrook and Cohn 7H10 agar (+OADC) with cycloheximide (500 microg/ml). Mycobacteria were recovered from five (71%) of seven pools. Detection of mycobacteria depended on the method used. High isolation rates (36-46% of the samples) were obtained by methods (A), (B) and (D). Contamination was a problem only with method (A). Inactivation of chlorine had a variable impact on mycobacterial detection. Isolates included M. kansasii, M. gordonae, M. fortuitum complex, M. sphagni, and M. vaccae, as well as M. simiae-like and M. chubuense-like organisms. In addition, a group of slowly growing and a group of rapidly growing isolates with previously unknown fatty acid and alcohol composition were isolated. No M. avium was detected. Mycobacterial counts were highest in a small pool with high temperature, low pH, and low content of free available chlorine.

Mycobacterium malmoense-specific nested PCR based on a conserved sequence detected in random amplified polymorphic DNA fingerprints.

Mycobacterium malmoense is an opportunistic human pathogen of increasing clinical importance. Since it is difficult to detect and identify the organism by conventional techniques, it was decided to seek a nucleic acid amplification method specific for M. malmoense. The method was based on detection of a conserved band in random amplified polymorphic DNA (RAPD) fingerprints of 45 M. malmoense strains. This band was a 1,046-bp product which was proven to be M. malmoense specific in dot blot hybridization analysis with a panel of mycobacterial strains belonging to 39 other species. The fragment was sequenced, and oligonucleotide primers were synthesized to evaluate the specificity of the PCR. Two primer pairs were found to be specific and sensitive in the nested PCR that was developed. All 49 M. malmoense strains analyzed produced a PCR product of the expected size. In contrast, no strains belonging to the other mycobacterial species tested produced amplicons with these primers under specified reaction conditions. The results of the electrophoresis were confirmed by the hybridization with the M. malmoense-specific oligonucleotide probe. This method could be applied to the analysis of clinical or environmental samples, permitting the rapid detection of M. malmoense.