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In the developed societies, day-care centers (DCCs) play an important role in the spread of antibiotic-resistant pneumococci both within the facility and from the facility to the community. This study was conducted to determine the effect of a nonavalent pneumococcal conjugate vaccine (PCV-9) on the carriage of antibiotic-resistant pneumococci in the DCC.Healthy DCC attendees ages 12 to 35 months were randomized to receive either PCV-9 or a control vaccine (conjugate meningococcus C vaccine) in a double blinded manner. Nasopharyngeal Streptococcus pneumoniae cultures were obtained from each subject before vaccination, monthly during the first year of follow-up and every 2 to 3 months during the second year of follow-up. For each isolate the serotype and antibiotic susceptibility were determinedA total of 132 and 130 evaluable toddlers received either PCV-9 or the control vaccine, respectively. In total 3748 cultures were obtained, of which 2450 (65%) were positive for S. pneumoniae. The resistance rates to penicillin, trimethoprim-sulfamethoxazole and erythromycin were 36, 35 and 16%, respectively. Resistance rates to ≥1 and ≥3 antibiotic categories were 52 and 9%, respectively. Antibiotic resistance was found mainly in the 5 serotypes included in the pneumococcal conjugate vaccines (6B, 9V, 14, 19F and 23F) and in 2 related serotypes (6A and 19A). In the vaccinated group a clear and significant reduction of the carriage rate of the serotypes included in the vaccine and the related serotype 6A as well as an increase in the carriage rate of the serotypes not included in the vaccine were observed. In parallel a significant decrease in carriage rate of antibiotic-resistant pneumococci was observed. The reduction of carriage of antibiotic-resistant pneumococci was seen in all age windows but was greater in the age window <36 months.The carriage rate of antibiotic-resistant S. pneumoniae, including multiply resistant S. pneumoniae, in DCC attendees is high. Pneumococcal conjugate vaccines seem to be an important tool for reducing the carriage rate of antibiotic-resistant pneumonia in DCCs.