Api strep

The material to microbiological tests included nasal and pharyngeal swabs collected from patients before a planned vaccination against S. pneumoniae (0m-1st test) and 1-2 months after vaccination (Im-2nd test). The swabs were collected using a kit with a transport medium, and next a culture of microorganisms was performed on the following media: Columbia agar with 5% sheep blood, Mannitol salt agar, McConkey, Chocolate agar, and Sabouraud (bioMerieux, France), and the phenotypic identification with kits: the APINH, the APIStrep, the APIE, the APINE APIStaph, and the API AUX (bioMerieux, France). The numeric code of the identified strain was read out with the apiweb™ program (bioMerieux, France). The tests were conducted according to the routinely applied microbiological diagnostics.

Milk samples were transferred with a calibrated loop (0.01 ml) onto Columbia agar (Oxoid, Great Britain) and Edwards medium (Oxoid, Great Britain), both supplemented with 5% defibrinated sheep blood. The plates were incubated at 37 °C for 48 ± 2 h under aerobic conditions. The grown cultures were examined microscopically after Gram staining, and the phenotypic traits were analysed (i.e., type of haemolysis, esculin hydrolysis, catalase production, CAMP reaction). For further analysis, only G+, catalase-negative cocci were selected. The final identification was performed using the commercial latex agglutination test Streptococcal Grouping Kit (Oxoid, Great Britain), API strep (bioMérieux, France) and PCR. All strains used in this study are summarized in Table 1.Table 1

Strains used in this study

Species	n
S.uberis	53
S. dysgalactiae	41
S. agalactiae	27
Other Streptococci	14

Clean catch mid-stream urine samples were collected from all participants using a wide-mouthed sterile-capped container. The specimen was promptly transported to the microbiology laboratory and cultured within one hour of collection. We used a 0.001 ml calibrated wire loop to inoculate samples onto Cystine Lactose Electrolyte Deficient (CLED) agar, blood agar, and chocolate agar (plates (Oxoid Ltd., Hampshire, United Kingdom) and incubate at 37°C for 24–48 hours. Colony counts yielding bacterial growth of 10⁵/ml of urine were regarded as significant for bacteriuria. Colony characteristics, Gram reaction, and biochemical reactions were used to identify bacterial isolates [29 (link), 30 ]. For the identification of Gram-negative bacteria, oxidase test, lactose fermentation, and hydrogen sulfur (H₂S) production in Kligler's iron agar (KIA) test, urease test, citrate test, and indole test were used. Gram positives were also identified using a type of haemolysis, a catalase test, and a coagulase test. Isolated bacteria were later confirmed using API 20E and API 20NE (bioMerieux) for Gram-negative bacteria and API-staph (bioMerieux) and API- Strep (bioMerieux) for Staphylococcus and Streptococcus species, respectively.