Cluster of Candida parapsilosis primary bloodstream infection in a neonatal intensive care unit

Cluster of Candida parapsilosis Primary Bloodstream Infection in a Neonatal Intensive Care Unit

Carmem Lúcia P. da Silva, Rosana M. R. dos Santos and Arnaldo Lopes Colombo

Disciplina de Doenças Infecciosas e Parasitárias, Depto. de Medicina Preventiva, UFRJ, Rio de Janeiro;Hospital da Lagoa, Rio de Janeiro; Disciplina de Doenças Infecciosas e Parasitaárias, Departamento de Medicina-UNIFESP, São Paulo, Brazil

Candida parapsilosis is an increasingly important bloodstream pathogen in neonatal intensive care units (NICU). We investigated a cluster of bloodstream infections in a NICU to determine whether nosocomial transmission occurred. During a 3-day period, 3 premature infants hospitalized in the same unit presented with sepsis caused by C. parapsilosis. Electrophoretic karyotype of the organisms was performed by using pulsed field gel electrophoresis in a countour-clamped homogeneous electric field system. The isolate from 1 newborn could not be typed, and the isolates from the remaining 2 infants had identical patterns. All 3 cases are described. We conclude that nosocomial transmission of C. parapsilosis occurred and that neonates under intensive care may represent a risk group for this pathogen.

Key Words: Candidemia, nosocomial infections, neonate.

The relative number of organisms causing nosocomial bloodstream infections has changed over the last decade, with Candida species now being firmly established as one of the most frequent agents worldwide [1, 2]. The number of nosocomial bloodstream infections due to Candida species in critically ill neonates is increasing. Taylor, et al., found that 21% of all cases of candidemia from a large institution, evaluated during a 7-year period, were documented in neonatal intensive care units (NICU) [3].

Nosocomial Candida infections have been recognized by many investigators as a major cause of morbidity and mortality in neonatal intensive care units [4, 5]. Newborns may acquire Candida species secondary to either vertical or horizontal transmission [6]. Risk factors identified by multivariate analysis of NICU populations include prior antibiotic exposure, hyperalimentation, the use of intravenous fat emulsions, and endotracheal intubation [7].

In the NICU setting, there are several reports of Candida albicans nosocomial cross infection [7-17].However, few studies using molecular DNA-based typing methods have demonstrated clusters of hematogenous candidiasis due to non-albicans species in such a population [15,18-20]. The present report describes 3 cases of C. parapsilosis candidemia documented during a 3-day period in a NICU in Rio de Janeiro, Brazil.

Materials and Methods

Between April 24 and 26, 3 cases of C. parapsilosis candidemia were documented in a NICU of a 159 bed private tertiary care hospital in Rio de Janeiro. All clinical and epidemiological data were retrospectively obtained by reviewing patients' clinical charts. In order to determine the background rate of C. parapsilosis bloodstream infections, the hospital's laboratory records were reviewed to identify all cases of positive blood cultures for C. parapsilosis reported from November 1, 1996, to May 31, 1997.

Cultures. Blood cultures of candidemic patients were obtained by venipuncture and incubated in a semi-automated Bactec System. The yeast isolates were identified at the species level by using the API-20C galleries (BioMerieux).

Susceptibility testing. Yeast profiles of susceptibility to amphotericin B, fluconazole and 5-flucytosine were performed by the NCCLS microdilution method [21].

Molecular Typing. In order to investigate the relatedness among C. parapsilosis isolates, we performed an electrophoretic karyotype (EK) of the organisms by using pulsed field gel electrophoresis in a countour-clamped homogeneous electric field system (CHEF-DRIII, Richmond-CA), following a previously described method [22, 23].

Briefly, colonies of C. parapsilosis were incubated overnight in 1% yeast extract, 2% peptone and 2% dextrose. After centrifugation, the cells were washed with 50 mM EDTA, pH 8.0 and incubated with lyticase. Agarose blocks were prepared by adding 1% low melt agarose to this solution. The blocks were incubated for 12 h in 0.01 M Tris buffer, 0.45M EDTA, 1% laurylsarcosine and 1mg of Proteinase K per mL. The washing procedures were repeated several times the next day and 2 mm of each agarose block were inserted in individual wells of an 8% chromosomic grade agarose gel. A countour-clamped homogeneous electric field was used to separate different molecular sizes of C. parapsilosis chromosomal DNA. Saccharomyces cerevisiae chromosome-DNA standard (BioRad) was inserted into the gel as a standard. In addition, 1 isolate of C. parapsilosis obtained from the blood culture of an adult patient hospitalized at another institution was included. The electrophoretic conditions were: 150 volts; 13°C switch time: 120 sec for 24 h and then 240 sec for 36 h. The gel was stained in ethidium bromide and phothographed. Two isolates were considered to have the same electrophoretic karyotype profile if all of the bands in one isolate matched the bands in another by visual comparison.

Results

According to data obtained by reviewing the laboratory records, only 3 cases of candidemia due to C. parapsilosis were documented in the NICU during the study period. No other candidemia was detected during the 6 months prior to the described outbreak The 3 candidemic patients were put side by side in the NICU; 2 of the patients were twin sisters.

Case 1 (Second Twin) April, 24. The neonate was delivered at 28 weeks gestation and weighed 770 g. At the age of 52 days, she developed a sepsis syndrome with blood and urine cultures positive for C. parapsilosis.

Prior to fungal infection: Mechanical ventilation and central venous catheter (CVC) were in place since birth. Phototherapy, hyperalimentation, and 38 days of antibiotics were also required.

Outcome: Candida parapsilosis was isolated from peripheral blood cultures and from the catheter tip that had been in place since day 34. Amphotericin B was started on day 54 at 1 mg/kg/day, and the CVC was removed on day 55. The last positive blood culture was obtained 3 days after the catheter removal. She received amphotericin B therapy for 18 days and was discharged at 136 days of age, 2 months after the end of treatment, without any sign of infection.

Case 2 (First Twin) April, 26. The neonate was delivered at 28 weeks gestation and weighed 910 g. At the age of 54 days, she developed a sepsis syndrome with blood cultures positive for C. parapsilosis.

Prior to fungal infection: Mechanical ventilation and CVC were in place for 42 days. Phototherapy, hyperalimentation, and 38 days of antibiotics were also required. Following case 1, there was a high suspicion for Candida infection and amphotericin B at 1 mg/kg/day was started at the very beginning of clinical signs of sepsis. No CVC had been in place 12 days prior to the occurrence of fungal sepsis.

Outcome: The patient received amphotericin B therapy for 18 days, and no additional positive culture was obtained. She was discharged at 127 days of age, almost 2 months after the end of treatment, without any sign of infection.

Case 3 April 26. The patient was delivered at 27 weeks gestation and weighed 700 g. At the age of 65 days, he developed a sepsis syndrome with blood and urine cultures positive for C. parapsilosis.

Prior to fungal infection: Mechanical ventilation and CVC were in place since birth. Phototherapy, parenteral nutrition and 46 days of antibiotics were also required. Outcome: Amphotericin B was started at 1 mg/kg/day and the central venous catheter was exchanged. Additional positive cultures for C. parapsilosis were obtained from tracheal aspirate on day 67, and blood cultures on days 69 and 77. Blood cultures remained positive up to the eleventh day of antifungal therapy, when the new CVC was removed. After that, all blood cultures were negative. Amphotericin B was maintained for a total of 24 days. The fungal infection was cured, but the patient died at 178 days of age due to bronchodysplasia complications.

Only 2 of the 3 C. parapsilosis isolates were available for further susceptibility testing and molecular typing. The amphotericin B, fluconazole and 5-flucytosine MIC values for both yeast isolates were 0.5 mg/mL, 0.25 mg/mL and 0.125 mg/mL, respectively. As illustrated in Figure 1, the electrophoretic karyotype of the 3 tested isolates exhibited 2 different patterns, one of them shared by both clinical isolates from the NICU patients, and the other exhibited by the reference strain.

Discussion

Apart from the increased risk for developing nosocomial infections, neonates admitted to a NICU are at an increased risk for infections due to unusual pathogens [24]. Knowledge of the epidemiology of nosocomial pathogens documented among newborns admitted to intensive care units is useful for clinicians to better treat this population and to anticipate possible threats. The emergence of Candida species as etiologic agents of nosocomial infections has been described [6]. In Brazil, recent data obtained from a multicenter study showed that 28 of 145 (19%) episodes of candidemia were reported among children admitted to the neonatal intensive care units of tertiary care hospitals [25].

Although Candida spp can be a perinatal pathogen, the predominant mode of acquisition in NICU populations is nosocomial transmission, which is even more important with C. parapsilosis infections [5, 6]. In Brazil, Levin, et al., have described a cluster of 6 cases of fungemia due to C. parapsilosis among adult patients [26]. To our knowledge, the present series represents the first report of a C. parapsilosis outbreak in a NICU setting described in this country.

The use of liquid glycerin given as a suppository was identified as a common source of a C. parapsilosis outbreak in a NICU [19]. In 2 other C. parapsilosis outbreaks, molecular based studies showed similar profiles among patients and hands of health care workers [20, 26]. Unfortunately, in the present report, we were not able to investigate the common source of the C. parapsilosis infections. However, the genomic relatedness of the 2 C. parapsilosis strains, and the geographic and temporal association are strong arguments for considering this occurrence a definite outbreak. We suggest that nosocomial acquisition through indirect patient contact could have occurred.

Very low birth weight (VLBW - birth weight less than 1,500 g) infants represent a special risk group of patients with increased susceptibility for fungal infections due to their impaired specific and nonspecific defense mechanisms [27]. Rates of fungal colonization among VLBW infants are over 20%, Candida albicans and C. parapsilosis being the most prevalent species; and 7.7% of the colonized patients later develop systemic Candida infections [8-16, 18-20, 28].The lower weight the baby, the higher the risk. In a prospective study, all infants who developed candidemia, weighed less than 1,000 g and were previously colonized [29]. In addition to baseline conditions, life saving and supporting measures such as broad spectrum antibiotics, mechanical ventilation, parenteral nutrition and central venous catheters are risk factors for fungal infection [7, 30, 31]. In our series, all 3 patients weighed less than 1,000 g at birth and were exposed to these risk factors previous to the fungal infection.

Among adult patients, systemic infections due to Candida parapsilosis have been associated with a lower mortality rate compared to other species of Candida 32-34]. This clinical finding is in accordance with experimental data showing that C. parapsilosis isolates exhibit a lesser potential for tissue invasion [35]. In the NICU population, mortality due to C. parapsilosis may be considerably higher, especially among the smallest babies [36]. In the largest outbreak of C. parapsilosis reported among neonates, 21 of 23 babies with systemic infection had a gestational age of less than 29 weeks, the case-fatality was 39%, and risk of death was 16-fold greater than that of controls [18].

The present cases of candidemia were successfully treated with amphotericin B. As expected, the 2 C. parapsilosis isolates were very susceptible to all of the antifungal drugs tested. Of interest, 1 patient continued to have positive blood cultures despite 11 days of high dose amphotericin B therapy. The cultures became negative just after the removal of the CVC catheter.

The issue of removing a central venous catheter in patients with candidemia is a matter of great controversy. In a recent report, we described a cohort of 145 patients with candidemia where catheter retention was recognized as an independent risk factor for death [32]. The case mentioned above illustrates that removing the catheter may influence clinical outcome.

Received on 14 July 2000; revised 3 November 2000.

Address for correspondence: Dr. Arnaldo Lopes Colombo, MD. Disciplina de Doenças Infecciosas e Parasitárias-UNIFESP. Rua Botucatu, 740. Zip Code: 04023-062. São Paulo-SP, Brazil. Fax/Phone: [phone]. Email: [e-mail]

• 1. Wright W.L., Wenzel R.P. Nosocomial Candida Epidemiology, transmission, and prevention. Infect Dis Clin North Am 1997;11:411-25.
• 2. Weinberger M., Sacks T., Sulkes J., et al. Increasing fungal isolation from clinical specimens: experience in a university hospital over a decade. J Hosp Infect 1997;35:185-95.
• 3. Taylor G.D., Buchanan Chell M., Kirkland T., et al. Trends and sources of nosocomial fungaemia. Mycoses 1994;37:187-90.
• 4. Khadilkar V., Tudehope D., Fraser S. A prospective study of nosocomial infection in a neonatal intensive care unit. J Paediatr Child Health 1995;31:387-91.
• 5. Reed S.E., Lasker B.A., Butcher D.S., et al. Nonperinatal nosocomial transmission of Candida albicans in a neonatal intensive care unit: prospective study. J Clin Microbio 1998;36:1255-9.
• 6. Waggoner Fountain L.A., Walker M.W., Hollis R.J., et al. Vertical and horizontal transmission of unique Candida species to premature newborns. Clin Infect Dis 1996;22:803-8.
• 7. Weese-Mayer D.E., Fondriest D.W., Brouillette R.T., Shulman S.T. Risk factors associated with candidemia in the neonatal intensive care unit: a case-control study. Pediatr Infect Dis J 1987;6:190-6.
• 8. Burnie J.P., Odds F.C., Webster C., Williams J.D. Outbreak of systemic Candida albicans in an intensive care unit caused by cross infection. Br Med J 1985;290:746-8.
• 9. Vaudry W.L., Tierney A.J., Wenman W.M. Investigation of a cluster of systemic Candida albicans infections in a neonatal intensive care unit. J Infect Dis 1988;158:1375-9.
• 10. Fraser C.A.M. Cross-infection and diversity of Candida albicans strain carriage in patients and nursing staff on an intensive care unit. J Vet Med Mycol 1990;28:317-25.
• 11. Sherertz R.J., Gledhill K.S., Hampton K.D., et al. Outbreak of Candida bloodstream infections associated with retrograde medication administration in a neonatal intensive care unit. J Pediatr 1992;120:455-61.
• 12. Betremieux P., Chevrier S., Quindos G., et al. Use of DNA fingerprinting and biotyping methods to study a Candida albicans outbreak in a neonatal intensive care unit. Pediatr Infect Dis J 1994,13:899-905.
• 13. Reagan D.R., Pfaller M.A., Hollis R.J., Wenzel R.P. Evidence of nosocomial spread of Candida albicans causing bloodstream infection in a neonatal intensive care unit. Diagn Microbiol Infect Dis 1995;21:191-4.
• 14. Faix R.G., Finkel R.D., Andersen R.D., Kostetter M.K. Genotypic analysis of a cluster of systemic Candida albicans infections in a neonatal intensive care unit. Pediatr Infect Dis J 1995;14:1063-8.
• 15. Vazquez J.A., Boikov D., Boikov S.G., Dajani A.S. Use of electrophoretic karyotyping in the evaluation of Candida infections in a neonatal intensive-care unit. Infect Control Hosp Epidemiol 1997;18:32-7.
• 16. Ruiz Diez B., Martinez V., Alvarez M., et al. Molecular tracking of Candida albicans in a neonatal intensive care unit: long-term colonizations versus catheter-related infections. J Clin Microbiol 1997;35:3032-6.
• 17. Fowler S.L., Rhoton B., Springer S.C., et al. Evidence for person-to-person transmission of Candida lusitaniae in a neonatal intensive-care unit. Infect Control Hosp Epidemiol 1998;19:343-5.
• 18. Saxen H., Virtanen M., Carlson P., et al. Neonatal Candida parapsilosis outbreak with a high case fatality rate. Pediatr Infect Dis J 1995;14:776-81.
• 19. Welbel S.F., McNeil M.M., Kuykendall R.J., et al. Candida parapsilosis bloodstream infections in neonatal intensive care unit patients: epidemiologic and laboratory confirmation of a common source outbreak. Pediatr Infect Dis J 1996;15:998-1002.
• 20. Huang Y.C., Lin T.Y., Leu H.S., et al. Outbreak of Candida parapsilosis fungemia in neonatal intensive care units: clinical implications and genotyping analysis. Infection 1999;27:97-102.
• 21. National Committee for Clinical Laboratory Standards. Reference method for broth dilution antifungal susceptibility testing of yeasts; Approved standard M27-Ş Wayne, Pa, National Committee for Clinical Laboratory Standards 1997;17(9):1-28.
• 22. Branchini M.L., Pfaller M.A., Rhine-Chalberg J., et al. Genotypic variation and slime production among blood and catheter isolates of Candida parapsilosis J. Clin. Microbiol 1994;32:452-6.
• 23. Colombo A.L., Branchini M.L., Geiger D., et al. Gastrointestinal translocation as a possible source of candidemia in an AIDS patient. Rev Inst Med Trop Săo Paulo 1996;38(3):197-200.
• 24. National Nosocomial Infections Surveillance-CDC. National Nosocomial Infections Report, Data Summary from October 1986-April 1996, AJIC 1996;24:380-8.
• 25. Colombo A.L., Nucci M., Salomăo R., et al. High rate of non-albicans candidemia in Brazilian tertiary care hospitals. Diagn Microbiol Infect Dis. 1999;34:281-6.
• 26. Levin A.S., Costa S.F., Mussi N.S., et al. Candida parapsilosis fungemia associated with implantable and semi-implantable central venous catheters and the hands of healthcare workers. Diagn Microbiol Infect Dis 1998;30:243-9.
• 27. Fleer A., Gerards L.J, Verhoef J. Host defense to bacterial infection in the neonate. J Hosp Infect 1988;11(Suppl A):320-7.
• 28. Baley J.E., Kliegman R.M., Boxerbaum B., Fanaroff A.A. Fungal colonization in the very low birth weight infant. Pediatrics 1986;78:225-32.
• 29. El-Mohandes A.E., Robbins L.J., Keiser J.F., et al. Incidence of Candida parapsilosis colonization in an intensive care nursery population and its association with invasive fungal disease. Pediatr Infect Dis J 1994;13:520-24.
• 30. Butler K.M., Baker C.J. Candida: An increasingly important pathogen in the nursery. Pediatr Clin North Am 1988;35:543-63.
• 31. MacDonald L., Baker C., Chenoweth C. Risk factors for candidemia in a children's hospital. Clin Infect Dis 1998;26:642-5.
• 32. Nucci M., Colombo A.L., Silveira F., et al. Risk factors for death in patients with candidemia. Infect Control Hosp Epidemiol 1998;19:846-50.
• 33. Meunier-Carpentier F. Fungemia in the immunocompromised host. Changing patterns, antigenemia, high mortality. Am J Med 1981;71:363-70.
• 34. Martino P., et al. Fungemia in patients with leukemia. Am J Med Sci 1993;306:225-32.
• 35. Weems J.J., Jr. Candida parapsilosis: epidemiology, pathogenicity, clinical manifestations, and antimicrobial susceptibility. Clin Infect Dis 1992;14:756-66.
• 36. Pacheco-Rios A., Avila-Figueroa C., Nobrigrot-Kleinman Santos J.I. Mortality associated with systemic candidiasis in children. Arch Med Res 1997;28:229-32.

Publication Dates

• Publication in this collection
  29 Oct 2001
• Date of issue
  Feb 2001

History

• Reviewed
  03 Nov 2000
• Received
  14 July 2000