An outbreak of nosocomial fungemia due to the unusual yeast, Pichia anomala occurred in the pediatric wards of our hospital over a period of 23 months (April 1996 to February 1998). A total of 379 neonates and children (4.2% admissions) were infected. The probable index case was admitted to the pediatric emergency ward, with subsequent transmission to the premature nursery, pediatric intensive care units, and other children wards. Carriage on the hands of health care personnel was likely to be responsible for dissemination of the fungus. The outbreak could only be controlled after a health education campaign to improve hand-washing practices was instituted and after nystatin-fluconazole prophylaxis to all premature neonates and high-risk infants was introduced. In a case-control study, we identified a lower gestational age, a very low birth weight (<1,500 g), and a longer duration of hospital stay as significant risk factors associated with P. anomala fungemia in premature neonates. We conducted a culture prevalence survey of 50 consecutive premature neonates and found that 28% were colonized with P. anomala at a skin or mucosal site on the date of delivery and that 20% of these neonates subsequently developed P. anomala fungemia. We performed multilocus enzyme electrophoresis on 40 P. anomala outbreak isolates (including patient and health care workers' hand isolates), and the results suggested that these isolates were identical. Our study highlights the importance of P. anomala as an emerging nosocomial fungal pathogen.Deep-seated fungal infections are important causes of morbidity and mortality in hospitalized patients (1, 2, 5, 13). Disseminated candidiasis is the most common nosocomial fungal infection, and Candida albicans has been reported to account for 50% to more than 70% cases of invasive candidiasis (2,5,6,8). However, recent reports have also suggested the emergence of infections caused by non-C. albicans candidas (3,14,21). In addition, less-common pathogenic yeasts (Malassezia, Trichosporon, Hansenula, and Rhodotorula spp.) have recently been reported, with increased frequency, as causes of nosocomial infections (7).Although a rare clinical isolate, the ascosporogenous yeast, Pichia anomala (formerly Hansenula anomala) has been implicated in causing fungemia in a neonatal intensive care unit (10), interstitial lung disease (19), endocarditis (12), and enteritis (9). In addition, there have been two reports of nosocomial outbreaks due to P. anomala: one in a Neonatal Intensive Care Unit in Liverpool, United Kingdom (10), and the other in an oncology hospital in Brazil (18). We describe here an outbreak of invasive P. anomala infection in the pediatric wards of our medical center that occurred during April 1996 to February 1998, with an attack rate of 4.2%. MATERIALS AND METHODSEpidemiologic investigation. The Nehru Hospital, affiliated with the Postgraduate Institute of Medical Education and Research, Chandigarh, India, is a 1,200-bed tertiary adult and pediatric referral center. The pediatric depa...

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Actions of salbutamol, disodium cromoglycate, and placebo administered as aerosols in acute asthma 725Resistance to DSCG in acute asthma is greater therefore, than to salbutamol. However, clinical observation suggests that resistance to beta-2 stimulants does also increase with severity of the attack. It would appear probable that only when severity exceeds the point at which measurements can be made with a peak flow meter does resistance to salbutamol become pronounced. Although placebo effects are greater in the attack than between attacks, again no information is available about attacks severe enough to prevent measurement using a peak flow meter. pipette into a capillary tube (for this technique blood pH capillary tubes cut to the same length as standard glass capillary tubes were used). They were sealed at one end by a flame and centrifuged for 15 minutes at 12 000 g, using a haematocrit centrifuge (Hawksley, London).After centrifugation the tubes were removed immediately and placed vertically. The length of the fat layer at the top and that of the solid layer at the bottom was measured with vernier callipers to the nearest 0 05 mm. Stool fat content was expressed as a percentage (steatocrit) of the total length of the solid column in the tube (that is fat layer + solid layer) (Fig. 1). The steatocrit was measured in duplicate. The stool fat content was also measured by Sobel's method2 which requires 3 g of stool.

bThe limitations of conventional methods of identification of Mycobacterium tuberculosis have led to the development of several nucleic acid amplification techniques which have the advantage of being rapid, sensitive, and specific. However, their expense or the need for technical expertise makes it difficult to use them in regions in which tuberculosis is endemic. A novel PCR restriction analysis (PRA) of the hsp65 gene was therefore developed for rapid screening of clinical isolates to identify Mycobacterium spp. The restriction enzymes NruI and BamHI were selected to obtain a limited number of restriction patterns to further differentiate between Mycobacterium tuberculosis complex (MTBC) and nontuberculous mycobacteria (NTM). Three hundred ten isolates from clinical specimens and 24 reference strains were tested. The assay correctly identified 295 of the 310 culture isolates as MTBC, while the remaining 15 isolates were identified as NTM. Of the isolates tested, 135 MTBC strains and all 15 NTM were also confirmed by PRA using Sau96I and CfoI. Thirty-eight randomly selected MTBC strains and all 15 NTM were further confirmed by sequencing. The NruI/BamHI PRA was simple, as it did not require any elaborate analyses. It was cost-effective, rapid, highly sensitive, and specific and did not require technical expertise. The assay can, therefore, be used as a simple screening test not only to detect Mycobacterium spp. but also to differentiate MTBC from NTM in peripheral laboratories with minimal availability of funds.

The high number of efflux genes overexpressed in an INH-resistant isolate with no known INH resistance associated mutations, suggests a role for efflux pumps in resistance to this antituberculous agent, with the role of Rv0194 and Rv0507 in INH resistance being reported for the first time.