Trends in serotype incidence and susceptibility (1997 to 2008) of Spanish Streptococcus pneumoniae pleural isolates (n ؍ 831) were explored. Penicillin (oral) nonsusceptibility rates and the incidence of 7-valent pneumococcal conjugate vaccine (PCV-7) serotypes showed decreasing trends (R 2 > 0.600; P < 0.002). In a recent report, years after licensure of the 7-valent pneumococcal conjugate vaccine (PCV-7), more than one-third of pediatric hospitalizations for pneumonia were complicated by empyema, with pediatric parapneumonic empyema a common cause of invasive pneumococcal disease (2). The use of PCV-7 has reduced the incidence of invasive disease in children (19), but annual empyema-associated hospitalization rates have increased (15).PCV-7 has been available in Spain since October 2001, only in the private market for healthy children, with a small impact on the incidence of pediatric parapneumonic empyema in Spain (16,17) given that vaccine coverage in 2006 was below 50% (13), and the disease detection has improved over time.Since rates of parapneumonic empyema have been increasing worldwide in the current decade (2-4, 7, 16, 17) despite PCV-7 introduction, it is essential to estimate the serotype epidemiology and resistance of Streptococcus pneumoniae isolates from pleural fluid. This study focuses on all isolates from pleural fluid received by the Spanish Reference Laboratory of Pneumococci (SRLP) from January 1997 to December 2008 through a passive, laboratory-based surveillance system. Isolates were serotyped by Quellung reaction and/or dot blot assay (11). Susceptibility was determined by agar dilution (12) according to CLSI (5) using Mueller-Hinton agar (Difco Laboratories, Detroit, MI) supplemented with 5% sheep blood (Biomedics, Madrid, Spain) and incubation under 5% CO 2 atmosphere. S. pneumoniae ATCC 6303 and S. pneumoniae ATCC 49619 plus five clinical isolates were used as quality controls (9). Current CLSI nonsusceptibility breakpoints for parenteral penicillin (Ն4 g/ml; nonmeningitis), oral penicillin (Ն0.12 g/ml), amoxicillin (Ն4 g/ml), cefotaxime (Ն2 g/ml; nonmeningitis), erythromycin (Ն0.5 g/ml), and levofloxacin (Ն4 g/ml) (6) were considered. Trends over time were explored by linear regression analysis. Tables 1 to 3 show yearly distribution of invasive and pleural isolates (with their susceptibilities) for the total population,