Blood agar with alpha hemolytic colony: Introduction, Possible pathogen and its details
5% sheep blood agar(BAP) is with growth of Streptococcus pneumoniae showing alpha (α) haemolytic colony as shown above picture. incomplete (α) hemolysis: Streptococcus pneumoniae, Streptococcus salivarius, viridans are referred to collectively as viridans streptococci, a name derived from viridis (Latin for “green”), referring to the green pigment formed by the partial, α-hemolysis of blood agar. Encapsulated, virulent strains of S. pneumoniae often forming highly mucoid, glistening colonies (production of capsular polysaccharide) surrounded by a zone of α -hemolysis. S. pneumoniae can also be isolated from above organisms being this sensitive towards the optochin and bile solubility test positive.
It is a Gram-positive elongated cocci ususally found in pairs i.e diplococci or lanceolate , alpha-hemolytic on blood agar facultative anaerobic , non-sporulated, non-motile but capsulated member of the genus Streptococcus. It resides asymptomatically in healthy carriers typically colonizing the respiratory tract, sinuses, and nasal cavity.
Pathogenicity of Pneumococcus
Pneumococcus is the most common cause of bloodstream infections, pneumonia, meningitis, and middle ear infections ( Otitis media) in young children. Severe infections can occur in the elderly and those already in poor health or immunosuppressed. Risk of infection is increased following splenectomy. In tropical and developing countries, S. pneumoniae is a major pathogen, responsible for acute life-threatening pneumonia and bacteraemia in those co-infected with HIV The invasive pneumococcal diseases include bronchitis, rhinitis, acute sinusitis, otitis media, conjunctivitis, meningitis, sepsis, osteomyelitis, septic arthritis, endocarditis, peritonitis, pericarditis, cellulitis, and brain abscess.Childhood pneumonia and serious infections are in patients with sickle cell disease.
Serotypes: Over 80 capsular serotypes where as less than 15 serotypes are responsible for most infections.
Laboratory Diagnosis of Pneumococcus
Specimens: Depending on the site of infection, specimens may include sputum, exudate, blood for culture, and cerebrospinal fluid.
Gram stain of specimen Gram positive elongated (lanceolate) diplococci and may show evidence of capsule but no evidence of spore.
Blood agar: Following overnight incubation. S. pneumoniae forms translucent or mucoid colonies, 1–2 mm in diameter. In young cultures the colonies are raised but later become flattened with raised edges, giving them a ringed appearance i.e ‘draughtsman’. Strains of some serotypes (e.g. serotype 3) produce large mucoid colonies. Pneumococci show alpha-haemolysis, i.e. colonies are surrounded by an area of partial haemolysis with a green-brown discoloration in the medium (reduced haemoglobin)
Optochin sensitivity Pneumococci are sensitive to optochin (ethylhydrocupreine hydrochloride). Placing a disc (5 µg) on a primary sputum culture and culturing the plate aerobically (not in CO2) can help to provide a rapid presumptive identification of pneumococcus.
Bile solubility test for S. pneumoniae
The bile (sodium deoxycholate) solubility test distinguishes S. pneumoniae from all other alpha-hemolytic (viridans) streptococci. It is bile soluble whereas all other alpha-hemolytic streptococci are bile resistant.
Agglutination test: A latex reagent to detect S. pneumoniae capsular antigen.
Useful antibacterial drugs are ampicillin, amoxycillin, erythromycin, co-trimoxazole, doxycycline, ofloxacin, vacomycin, chloramphenicol (nitrofurantoin in case of urine ), teicoplanin, linezolid. Penicillin-resistant strains are becoming an increasing problem.
Streptococcus pneumoniae shows alpha haemolysis on blood agar under aerobic conditions where as beta-hemolysis under anaerobic condition
Absence of capsules in Gram stain if organisms from cultures.
It also forms short chains in Gram stain particularly following culture but lacking such features in specimens.
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